Drug conjugates based on ε-poly-L-lysine, intermediates thereof and uses thereof

Abstract

The present invention discloses a drug conjugate based on ε-poly-L-lysine, its intermediate and its use. The present invention provides an ε-poly-L-lysine derivative-drug conjugate compound having a controllable number of group bonds and a controllable number of group bond sites as shown in formula (I). The drug conjugate of the present invention has one or more advantages of low renal clearance, low hepatic clearance and low splenic clearance, long plasma half-life, rapid accumulation of effective drug concentration in the lesion, and better therapeutic effect. [Case 1] TIFF2024542139000558.tif28169

Description

Detailed Description of the Invention

[0001] This application claims the priority of Chinese Patent Application No. 2021113237547 with an application date of November 8, 2021. This application incorporates the entire content of the said Chinese patent application by reference.

[0002] [Technical Field] The present invention relates to drug conjugates based on ε-poly-L-lysine, intermediates thereof, and their uses.

[0003] [Background Art] Chemotherapeutic drugs usually have drawbacks such as non-target organ toxicity and low tumor-specific accumulation. Therefore, a series of special delivery forms for chemotherapeutic drugs have been developed, including self-assembled delivery systems such as liposomes, micelles, vesicles, and microspheres, as well as drug conjugate systems such as low molecular weight prodrugs, high molecular weight drug conjugates, and antibody-drug conjugates. Among them, the following high molecular weight drug conjugates have been reported: a) Polyethylene glycol-block copolymer-drug conjugates, such as NC-6004 (PEG-b-poly(glutamic acid)-cisplatin) (British Journal of Cancer, 2011, 104, 593), K-912 / NC-6300 (polyethylene glycol-b-poly(glutamic acid)-epirubicin) (Investigational New Drugs, 2017, 35, 307), etc.; b) Core crosslinked: CriPec docetaxel (copolymeric micelles of docetaxel containing methacrylic acid moieties and PEG block polymers) (Biomaterials, 2015, 53, 370); c) 4-arm PEG drug conjugates, such as NKTR102 (4-arm polyethylene glycol-irinotecan coupling) (Polymers, 2014, 6, 2186; Lancet Oncology, 2015, 16, 1556; Mol Cancer Ther, 2018, 17, 196), DFP-13318 (4-arm polyethylene glycol-SN-38 conjugate) (Journal of Medicinal Chemistry, 2014, 57, 2303); d) Random drug conjugates with a linear polymer as the parent and reactive functional groups in the side chains, such as Opaxio (polyglutamic acid-paclitaxel conjugate) (British Journal of Cancer, 2008, 98, 1608); e) Random drug conjugates and pharmacokinetic regulators with a linear polymer as the parent and reactive functional groups in the side chains, such as OsteoDex (dextran grafted alendronic acid) (Int J Oncol, 2010, 37, 563; Anticancer Research, 2016, 36, 6499); f) CRLX101 (cyclodextrin polyethylene glycol copolymer, side chain bound CPT) (Clinical Cancer Research, 2015, 21, 808; J Control Release, 2011, 153, 49), CRLX301 (cyclodextrin polyethylene glycol copolymer, side chain bound docetaxel) (Journal of Clinical Oncology, 2016, 34(15_suppl), 2526); g) Highly branched polymer drug conjugates or dendrimer drug conjugates, such as the dendrimer poly-L-lysine drug conjugate from Starpharma (Mol Pharm, 2018, 15, 4568).

[0004] The structural formula of ε-poly-L-lysine hydrochloride is [C6H 12It is [N2O-HCl]n-H2O, and its use as a biopreservative has been approved by the Ministry of Health, Labour and Welfare of Japan, the FDA of the United States, South Korea, China, and many other countries and organizations. Currently, the forms of ε-poly-L-lysine for drug delivery include block copolymers (WO2019039544) and graft copolymers (Mol Pharm, 2015, 12, 4226; Adv Mater, 2016, 28, 8162; CN109265680A). Among them, the block copolymer form only utilizes the ε-poly-L-lysine terminal group functionality, with a low degree of modification and low adjustability; the ε-poly-L-lysine graft copolymer obtained by random grafting has an uncontrollable distribution of graft functional groups on the polymer backbone. As a result, chain-to-chain heterogeneity of the polymer occurs, and these ε-poly-L-lysine graft copolymers obtained by the degree and distribution of modification have different pharmacological and pharmacodynamic properties in vivo. Therefore, in the existing ε-poly-L-lysine random modification method, in addition to the molecular weight, molecular weight distribution of ε-poly-L-lysine, and average degree of modification of a specific coupling molecule, it is necessary to examine the modification distribution of the coupling molecule between different batches, making it difficult to set quality standards.

[0005] According to the published literature (Mol Pharm, 2018, 15, 4568), it is suggested that there is a significant correlation between the molecular size of dendritic polymer-drug conjugates and the uptake, retention, release, and clearance of drugs in lesions. When the size of the dendrimer is small, the drug is released more quickly but is easily excreted from the kidneys; increasing the number of synthetic generations of the dendrimer polymer to increase the size of the dendrimer increases the steric resistance of the outer layer space and is disadvantageous for drug release from the enzymatically cleaved linker. As a result, it becomes difficult to balance the relationship between the clearance, retention, and drug release of dendrimer-drug conjugates, and it is difficult to obtain an optimal therapeutic effect.

[0006] For example, the dendritic polylysine drug conjugate developed by Starpharma is synthesized by one-point divergent synthesis. Due to the dendritic polylysine, high surface functional group density, and large steric resistance between the drug and the dendritic polylysine body, the accessibility of enzymes to the conjugate between the drug and the dendritic polylysine body is highly correlated with the generation of the dendritic polymer and the surface PEG chain length, and it is necessary to improve it by reducing the molecular weight. To increase the drug release rate, it is necessary to improve it by reducing the generation number of the dendrimer or shortening the PEG chain length (Mol Pharm, 2018, 15, 4568). According to the literature, only the dendritic polylysine-drug conjugate with a small molecular weight (G4 PEG570 , a fourth-generation dendrimer with an average molecular weight of about 2.26×10 4 and a coupling polyethylene glycol with an average molecular weight of 570) could achieve a faster drug release rate (48% release in 48 hours). When the PEG chain length was increased (G4 PEG1100 , a fourth-generation dendrimer with an average molecular weight of about 2.86×10 4 and a coupling polyethylene glycol with an average molecular weight of 1100), or when the generation number of the dendritic polylysine was increased (G5 PEG570 , a fifth-generation dendrimer with an average molecular weight of about 4.51×10 4 and a coupling polyethylene glycol with an average molecular weight of 570), the drug release rate decreased (29% release rate in 48 hours). When both the generation number of the dendritic polylysine and the chain length of polyethylene glycol were increased (G5 PEG1100 , a fifth-generation dendrimer, an average molecular weight of about 6.11×10 4 , and a coupling polyethylene glycol with an average molecular weight of 1100), the drug release rate decreased dramatically, and only 5% of the drug was released within 48 hours of administration). Although the drug release rate can be increased by reducing the size of the dendrimer, the problem of renal clearance will be faced again. According to the literature (Mol Pharm 2008, 5, 449), the specific renal clearances of different dendritic polylysines in 0-24 hours were as follows: Lys 16 (PEG 570 ) 32 was 40.9% (average molecular weight 2.24×10 4 ), Lys8(PEG2000 ) 16 at 6.6% (average molecular weight 3.41×10 4 ), Lys 16 (PEG 2000 ) 32 at 0.6% (average molecular weight 6.8×10 4 ). From this, in order to minimize the renal clearance of dendritic polylysine, it is necessary to couple a higher molecular weight PEG (for example, PEG 2000 ) to obtain a polymer with a total molecular weight exceeding 60,000. It was found that a higher molecular weight dendritic polymer and long PEG modification on the surface mean higher steric hindrance, leading to a decrease in drug release rate and efficacy. Therefore, in current dendrimers as drug delivery systems, only the in vivo clearance rate of the polymer and the drug release rate within the tumor can be balanced without achieving an optimal effect.

[0007] [Summary of the Invention] The technical problem to be solved by the present invention is directed to the drawback that in prior art dendrimer drug conjugates, it is difficult to balance the relationship among clearance, retention, and drug release, and it is difficult to obtain an optimal therapeutic effect. Therefore, a drug conjugate based on ε-poly-L-lysine, an intermediate thereof, and its use are provided. The drug conjugate of the present invention has one or more of the advantageous effects of low renal clearance, low hepatic clearance, and low splenic clearance, a long plasma half-life, rapid accumulation of effective drug concentration at the lesion site, and a better therapeutic effect.

[0008] The object of the present invention is to provide an ε-poly-L-lysine derivative-drug conjugate having a controllable number of group couplings and controllable number of group coupling sites as shown by formula (I):

[0009] [Chemical Formula]

[0010] The ε-poly-L-lysine derivative-drug conjugate comprises the following: Containing an ε-poly-L-lysine residue represented by formula (II), wherein the number of repeating units of the ε-poly-L-lysine moiety is n

[0011]

Chemical formula

[0012] Here, the ε-poly-L-lysine is of biosynthetic origin or chemical synthetic origin; The value of n is selected from integers of 4 to 100; Y is at least a trifunctional branching center; P is a pharmacokinetic regulator residue; D is a pharmacologically active agent residue; X is the terminal group X; L 0 、L 1 、L 2 are each independently a covalent bond or a C1-C linker containing or not containing heteroatoms, where the heteroatom is O, S, Se, N, P, Si, or B, and the number of said heteroatoms is one or more. In the case of one or more, the heteroatoms are the same or different; the linker contains or does not contain an unsaturated group; the L 40 connects the ε-poly-L-lysine residue and Y, the L 0 connects P and Y, and the L 1 connects D and Y. 2 connects D and Y.

[0013] In some embodiments, the value of the number n of repeating units of the ε-poly-L-lysine moiety is selected from integers of 4 to 100.

[0014] In some embodiments, the value of the number n of repeating units of the ε-poly-L-lysine moiety is selected from integers of 10 to 70.

[0015] In some embodiments, the value of the number n of repeating units of the ε-poly-L-lysine moiety is selected from integers of 20 to 40; for example, selected from 30.

[0016] In the ε-poly-L-lysine derivative-drug conjugate of the present invention, Y is at least three functional branching centers having the following structures, or a polyfunctional branching center composed of two or more of the following structures:

[0017]

Chemical formula

[0018] Here, Z is O, S, S(O), S(O)2, NR a , CHR 0 ; R 0 is H, D, halogen, nitro group n, cyano group, C1-C 10 alkyl group, C1-C 10 alkoxy group, C3-C 10 alkenyl group, C3-C 10 alkynyl group, C3-C8 cycloalkyl group, C2-C8 heterocycloalkyl group, C6-C 10 aryl group, C5-C 10 heteroaryl group, or selected from primary amine, secondary amine, tertiary amine, hydroxyl, mercapto acid, carboxylic acid, ester, amide, boronic acid, boronic acid ester, phosphoric acid, sulfonic acid, sulfoxide, aldehyde, ketone functional group; the heteroatom in the C2-C8 heterocycloalkyl is O, S or N, the number of the heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different; the heteroatom in the C5-C 10 heteroaryl is O, S or N, the number of the heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different; R a is H, C1-C 10 alkyl group, C1-C 10 alkoxy group, C3-C 10 alkenyl group, C3-C 10 alkynyl group, C3-C8 cycloalkyl group, C2-C8 heterocycloalkyl group, C6-C 10 aryl group or C5-C10 selected from heteroaryl groups; the heteroatoms in the C2-C8 heterocycloalkyl group are O, S or N, the number of said heteroatoms is one or more, and when there are one or more, said heteroatoms are the same or different; the C5-C 10 the heteroatoms in the heteroaryl group are O, S or N, the number of said heteroatoms is one or more, and when there are one or more, said heteroatoms are the same or different; Ar is C6-C 20 aryl or C5-C 20 heteroaryl, and the heteroatoms in the C5-C 20 in the heteroaryl are O, S or N: the number of said heteroatoms is one or more, and when there are one or more, said heteroatoms are the same or different; b is an integer from 0 to 3.

[0019] In some embodiments, the branching center Y is a substituted or unsubstituted amino acid (residue) having a functionality of at least 3, said amino acid being a natural amino acid or a non-natural amino acid, said amino acid having an arrangement of D-type, L-type, or a mixture of D / L-type, and when said amino acid is a mixture of D / L-type, the L-type arrangement has a proportion greater than 0% and less than 100%.

[0020] In some embodiments, the branching center Y is an amino acid (residue) having a functionality of 3, and is selected from one or more of aspartic acid, glutamic acid, lysine, ornithine, arginine, citrulline, histidine, serine, threonine, tryptophan, tyrosine, hydroxyproline, cystine, or cysteine: the stereoconfiguration of said amino acid is D-type or L-type or a mixture of D / L-type, and when said amino acid is a mixture of D / L-type, the L-type occupies 0% to 100%.

[0021] In some embodiments, the linker L 0 and L 1 are each independently a covalently bonded, environmentally responsive linker, or a non-environmentally responsive linker, and L 2 is the structure L2a -L 2b is an environmentally responsive linker, where L 2a is linked to Y, and L 2b is linked to D in a D-type manner; L 2a and L 2b are each independently a covalent bond or an environmentally responsive linker; the structure of the ε-poly-L-lysine derivative-drug conjugate is as shown in formula (III):

[0022]

Chemical formula

[0023] where X, Y, P, D, L 0 , L 1 are each defined as described in compound (I).

[0024] In some embodiments, the linker L 0 and L 1 are covalently bonded, L 2 is an environmentally responsive linker having the structure L 2a -L 2b where L 2a is linked to Y, and L 2b is linked to D in a D-type manner; L 2a and L 2b are each independently a covalent bond or an environmentally responsive linker; the structure of the ε-poly-L-lysine derivative-drug conjugate is as shown in formula (IV):

[0025]

Chemical formula

[0026] where X, Y, P, and D are each defined as described in compound (I).

[0027] In some embodiments, when the electrophilic group of the Y molecule is bonded to L 2 , L 2 =L 2bWhen L 2a does not exist, the linker L 0 of the trifunctional branching center Y 1 and L 2 is selected from any of the following structures:

[0028]

Chemical formula

[0029] ; preferably

[0030]

Chemical formula

[0031] More preferably

[0032]

Chemical formula

[0033] .

[0034] In some embodiments, when the nucleophilic group of the Y molecule is bonded alone to L 2 or L 2a or together with L 2a and L 2b , in which case L 2 =L 2a or L 2 =L 2a -L 2b and the linker L 0 of the trifunctional branching center Y 1 and L 2 is selected from any of the following structures:

[0035]

Chemical formula

[0036] ; preferably

[0037]

Chem.

[0038] 。

[0039] In some embodiments, the terminal group X is OR, SR, NR 1 R 2 , a carboxy protecting group, or L 2b -D, and is selected from the group consisting of: said R, R 1 , R 2 are independently selected from H, C1-C 30 alkyl, C1-C 30 alkoxy, C3-C 30 alkenyl, C3-C 30 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C 20 aryl, or C5-C 20 heteroaryl; R 1 , R 2 and the linked N atom may form a C2-C8 heterocycloalkyl; the heteroatom in the C2-C8 heterocycloalkyl is O, S or N, the number of heteroatoms is one or more, and when there are one or more, the heteroatoms are the same or different; the C5-C 20 heteroaryl is O, S or N, the number of heteroatoms is one or more, and when there are one or more, the heteroatoms are the same or different.

[0040] In some embodiments, the terminal group X is OR, SR, NR 1 R 2 , a carboxy protecting group, or L 2bSelected from -D, said R, R 1 , R 2 is H, C1 - C 10 alkyl, or R 1-1 substituted C1 - C 10 alkyl, independently selected; said R 1-1 is -NH2 or

[0041]

Chemical formula

[0042] is.

[0043] In some embodiments, the environment-responsive linker is one or more of an enzyme-responsive linker, a pH-responsive linker, a light-responsive linker, and a redox-responsive linker.

[0044] In some embodiments, the enzyme-responsive linker is cleavable by one or more of the following enzymes: secreted phospholipase A2, acid phosphatase, serum alkaline phosphatase, cytochrome P450, sulfatase, prostate-specific antigen, phospholipase A1, phospholipase A2, phospholipase B, phosphodiesterase C, phosphodiesterase D type, neutrophil elastase, cysteine protease-3, histone protease, matrix metalloprotease, β-glucosidase, β-galactosidase, DTP, nitroreductase, reduced coenzyme II, aminopeptidase N, carboxylesterase, myocardial xanthine oxidase, histone deacetylase, asparagin endopeptidase, urokinase-type fibrinogen activator, urokinase-type fibrinogen activator receptor, collagenase.

[0045] In some embodiments, the enzyme-responsive linker is cleavable by one or more of the following enzymes: cysteine protease-3, histone protease, matrix metalloprotease, β-glucosidase.

[0046] In some embodiments, the enzyme-responsive linker comprises one of the following amino acid sequences (residues): Cit-Phe, Lys-Lys, Phe-Lys, Arg-Arg, Val-Cit, Val-Ala, Val-Lys, Ala-Lys, Phe-Cit, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Arg, Asn-Pro-Val, Gly-Pro-Nle, Glu-Val-Cit, Glu-Val-Ala, Gly-Phe-Gly, Gly-Phe-Phe, Gly-Leu-Gly, Gly-Val-Ala, Gly-Phe-Leu-Gly, Gly-Phe-Phe-Leu, Gly-Leu-Leu-Gly, Ciy-Phe-Tyr-Ala, Gly-Phe-Gly-Phe, Ala-Gly-Val-Phe, Gly-Phe-Phe-Gly, Gly-Gly-Phe-Gly, Asp-Glu-Val-Asp, Gly-Phe-Leu-Gly-Phe, Gly-Phe-Ala-Gly-Leu-Phe, Gly-Leu-Ala-Ala-Val-Ala, Gly-Gly-Phe-Leu-Gly-Phe or Gln-Ser-Phe-Arg-Phe-Lys.

[0047] In some embodiments, the enzyme-responsive linker comprises the following structure:

[0048] [Chemical formula] TIFF0007693948000017.tif254169TIFF0007693948000018.tif135169

[0049] .

[0050] In some embodiments, the pH-responsive linker comprises one or more of the following structures: hydrazone, imine, oxime, carboxylic acid ester, thioester, sulfate ester, sulfonate ester, protonate, carbonate ester, carbamate, substituted carbamate, ketone acetal, acetal, silyl ether, phosphate ester, borate ester, phosphoramidite, or cis-aconitic acid group.

[0051] In some embodiments, the pH-responsive linker comprises the following structure:

[0052] [Chemical formula]

[0053] Here, m = 0 to 4.

[0054] In some embodiments, the photo-responsive linker comprises one or more of the following structures: o-nitrobenzene, coumarin, benzoin, BODIPY, or cyanine group.

[0055] In some embodiments, the photo-responsive linker comprises the following structure:

[0056] [Chemical formula]

[0057] In some embodiments, the redox-responsive linker comprises one or more of the following structures: thioketone, phenyl borate, phenylboronic acid, oxalate, vinyl ether, thioether group, aminoacrylate, disulfide, diselenide, 2,4-dinitrobenzenesulfonate, 2-azidomethyl benzoate, 4-azidobenzyl, unsaturated acid ester, or azobenzene group.

[0058] In some embodiments, the redox-responsive linker comprises the following structure:

[0059] [Chemical formula]

[0060] In some embodiments, linker L 2a is represented by formula (V):

[0061] [Chemical formula]

[0062] Said Q is

[0063] [Chemical formula]

[0064] selected from one of; Here, R 3 and R 4 are independently selected from H, D, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 alkenyl, C3-C6 alkynyl, C3-C8 cycloalkyl, C2-C8 heterocycloalkyl, C6-C 10 aryl, or C5-C 10 heteroaryl; R 3 , R 4 and the linked C atoms may form C3-C8 alkyl or heterocycloalkyl; the heteroatom in the C2-C8 heterocycloalkyl is O, S or N, and the number of the heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different; the C5-C 10 heteroaryl is O, S or N, and the number of the heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different; W is a covalent bond, or a C0-C 20 fragment containing or not containing heteroatoms, and the heteroatom is O, S, Se, N, P, Si, or B; here, the number of heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different; the W fragment may contain a bond not containing heteroatoms or a bond not containing unsaturated bonds; Z is

[0065] [Chemical formula]

[0066] Selected from one of them, R a is defined as described in the definition at the branching center Y.

[0067] In some embodiments, the linker L 2a is selected from the following structures or covalent bonds:

[0068]

Chemical formula

[0069] Here, A is O, S, S(O), S(O)2, NR a , C(R 3 R 4 ); p is selected from integers from 0 to 16; q is selected from integers from 0 to 16; m is selected from integers from 0 to 4; R a is as described in the definition at the branching center Y; R 3 and R 4 are as described in the definition in formula (V); Said L 2a is preferably the following structure:

[0070]

Chemical formula

[0071] L 2a -L 2b -D, in L 2a when is

[0072]

Chemical formula

[0073] is the case, the divalent aryl group is via a carbon atom to L 2b or D (L 2bBy linking in the case of, for example,

[0074]

Chemical formula

[0075] rings such as are formed.

[0076] In some embodiments, the linker L 2b is selected from the following structures:

[0077]

Chemical formula

[0078] .

[0079] In some embodiments, the linker L 2b is selected from the following structures:

[0080]

Chemical formula

[0081] L 2b is

[0082]

Chemical formula

[0083] in the case of,

[0084]

Chemical formula

[0085] is linked to D via an oxygen atom,

[0086]

Chemical formula

[0087] form a ring such as

[0088] In some embodiments,

[0089]

Chem.

[0090] is

[0091]

Chem.

[0092] and, for example,

[0093]

Chem.

[0094] is.

[0095] In some embodiments, the pharmacokinetic modulator residue P is selected from a polyethylene glycol derivative residue having a repeat unit number a and a terminal group R b , a hyaluronic acid derivative residue having a repeat unit number b, a polyphosphate ester residue having a repeat unit number c, a polyinosine residue having a repeat unit number d, or a polyoxazoline residue having a repeat unit number f; where R b is H, an alkyl group of C1-C 10 , a heteroalkyl group of C1-C 10 , a cycloalkyl group of C3-C 10 , an alkenyl group of C3-C 10 , or an alkenyl group of C3-C 10is an alkynyl group or a hydroxy protecting group; a is selected from integers of 5 to 250; b is selected from integers of 5 to 250; c is selected from integers of 5 to 250; d is selected from integers of 5 to 250; e is selected from integers of 5 to 250; f is selected from integers of 5 to 250.

[0096] In some embodiments, the pharmacokinetic modulator residue P is a polyethylene glycol derivative residue having a repeating unit number a and a terminal group R b where a is selected from integers of 5 to 150 and b is selected from integers of 5 to 150.

[0097] In some embodiments, the pharmacokinetic modulator residue P is a polyethylene glycol derivative residue having a repeating unit number a and a terminal group R b where a is selected from integers of 5 to 150, preferably integers of 10 to 60, more preferably integers of 15 to 50, such as 21, 43 or 44. In some embodiments, the pharmacokinetic modulator residue P is a residue of a polyethylene glycol derivative having a repeating unit number a and a terminal group R b where R b is an alkyl group of C1-C 10 , preferably an alkyl group of C1-C6, more preferably an alkyl group of C1-C3, such as a methyl group, an ethyl group, n-propyl or isopropyl.

[0098] In some specific embodiments, the pharmacokinetic modulator residue P is a methyl-capped polyethylene glycol derivative residue having a number average molecular weight of about 550, 1,000, 2,000, 3,000, 4,000, or 5,000.

[0099] In some specific embodiments, the pharmacokinetic modulator residue P is a methyl-capped polyethylene glycol derivative residue having a number average molecular weight of about 1,000 or 2,000.

[0100] In some embodiments, the polyethylene glycol derivative residue is selected from the following structures:

[0101] [Chemical formula]

[0102] Here, a is selected from integers from 5 to 150, b is selected from integers from 5 to 150, and r is selected from integers from 0 to 8.

[0103] In some embodiments, the polyethylene glycol derivative residue is

[0104] [Chemical formula]

[0105] wherein a is selected from integers from 20 to 45, b is selected from integers from 5 to 10, and r is selected from integers from 0 to 3.

[0106] In some embodiments, the polyethylene glycol derivative residue is

[0107] [Chemical formula]

[0108] is.

[0109] In some embodiments, the pharmaceutical active agent D in the ε-poly-L-lysine derivative-drug conjugate may have an active functional group selected from one or more of primary amine, secondary amine, tertiary amine, hydroxyl, mercapto, carboxyl, ester, amide, boronic acid, boronate, phosphoric acid, sulfonic acid, sulfoxide, aldehyde, and ketone groups.

[0110] The pharmaceutically active agent is selected from one or more of an anesthetic, an antacid, an anti-infective agent, a cardiovascular agent, a diuretic, a blood tonic, an immunosuppressive agent, a GLP-1 agonist, a hormone and analogs, an ophthalmic agent, a pain therapeutic agent, a respiratory agent, an anti-arthritis agent, an anticonvulsant, an antihistamine, an anti-inflammatory agent, an anti-ulcerative disease agent, a behavior-modifying agent, an anti-tumor agent, an anti-cancer antigen, a central nervous system agent, a psychiatric therapeutic agent, a contraceptive, a diabetes therapeutic agent, a growth promoter, a hemostatic agent, an immunostimulant, an immunomodulator, a muscle relaxant, an obesity therapeutic agent, an osteoporosis therapeutic agent, a tranquilizer, a urinary acidifier, a vitamin agent, a peptide agent, an oligonucleotide agent, an mRNA agent, an antibody agent, a biologic, a target proteolytic agent, a PROTAC agent, an oligosaccharide agent or a target agent.

[0111] In particular, the pharmaceutically active agent residue D is an anti-malignant tumor agent residue, and the anti-malignant tumor agent includes one or more of a tumor target agent, a PROTAC agent, a targeted proteolytic agent, a tumor immunomodulator or a chemotherapeutic agent.

[0112] Anticancer drugs include abemaciclib, abiraterone, abrocitinib, acalabrutinib, afatinib, aldesleukin, alectinib, alflutinib, alemtuzumab, altretamine, amcenestrant, aminoglutethimide, amsacrine, anastrozole, anlotinib, apalutamide, apatinib, arzoxifene, asimicinib, asparaginase, avapritinib, avitinib, axitinib, azacitidine, baricitinib, belinostat, bendamustine, bexarotene, bicalutamide, biciromol, binimetinib, bleomycin, boanamycin, bortezomib, bosutinib, brigatinib, buserelin, busulfan, cabazitaxel, cabozantinib, carfilzomib, carmustine, carmofur, cedazuridine, ceritinib, cetrorelix, chidamide, chlorambucil, cisplatin, cladribine, clofarabine, cobimetinib, colchicine, copanlisib, crizotinib, cyclophosphamide, cytarabine, dabrafenib, dacarbazine, dacomitinib, dactinomycin, darolutamide, dasatinib, daunorubicin, decitabine, degarelix, delgocitinib, denileukin, deruxtecan, docetaxel, donafenib, doxorubicin, duvelisib, enasidenib, encorafenib, ensartinib, entrectinib, enzalutamide, enzastaurin, erastin, epirubicin, erdafitinib, eribulin, erlotinib, estradiol, estramustine, etoposide, everolimus, exemestane, fasudil, fedratinib, filgotinib, flucytosine, fludarabine, fulvestrant, fluvastatin, formestane, fostamatinib, fruquintinib, fulvestrant, gefitinib, gemcitabine, gilteritinib, giredestrant, glasdegib, goserelin, histrelin, hydroxyurea, ibrutinib, ibudilast, icariin, icotinib, idarubicin, idelalisib, ifosfamide, imatinib, imiquimod, infliximab, ingenol mebutate, interferon alpha-2b, irinotecan, ibosidenib,Ixabepilone, Ixazomib, Lanreotide, Lapatinib, Larotrectinib, Lenalidomide, Lenvatinib, Letrozole, Leucovorin, Leuprolide, Lomustine, Lonafarnib, Lorlatinib, Lurbinectedin, Maytansine, Mechlorethamine, Medroxyprogesterone, Megestrol, Melphalan, Melphalan Flufenamide, Mercaptopurine, Methotrexate, Methoxsalen, Methylprednisolone, Midostaurin, Mitomycin, Mitotane, Mitoxantrone, Mitozolomide, Mobocertinib, Monomethylauristatin E, Monomethylauristatin F, Nelarabine, Nandrolone, Neratinib, Neralizumab, Nilotinib, Nilutamide, Nintedanib, Niraparib, Octreotide, Olaparib, Olmutinib, Omacetaxine, Orelabrutinib, Osimertinib, Oxaliplatin, Paclitaxel, Pacritinib, Palbociclib, Pamidronate, Pamiparib, Panobinostat, Pazopanib, Pemigatinib, Pegaptanib, Pegaspargase, Peginterferon Alfa-2b, Pemigatinib, Pemetrexed, Pentetreotide, Pentostatin, Pexidartinib, Phenoxybenzamine, Pidotimod, Purinethol, Pritidepsin, Pomalidomide, Ponatinib, Porfimer, Pralatrexate, Pralsetinib, Prednisolone, Procarbazine, Pyrotinib, Qizaltinib, Radotinib, Raloxifene, Raltitrexed, Regorafenib, Ribociclib, Rintatolimod, Repotrectinib, Romidepsin, Rubraca, Luxolitinib, Sabatolimab, Selinexor, Selpercatinib, Sermetinib, Sonidegib, Sorafenib, Sotorasib, Streptozocin, Sunitinib, Sulfatinib, Talazoparib, Tamoxifen, Tazemetostat, Tegafur, Temozolomide, Temsirolimus, Teniposide, Tepotinib, Teprenone, Thalidomide, Thioguanine, Thiotepa, Thyrotropin Alfa, Tipiracil, Tipifarnib, Tirbanibulin, Tiboxanib, Trametinib, Tofacitinib, Topotecan, Toremifene, Trabectedin, Tretinoin, Trifluridine, Trilaciclib, Triptorelin, Tsukatinib, Upadacitinib, Umbralisib, Ucedrone, Uroacitide, Valbicin, Vandetanib,Selected from one or more of vemurafenib, venetoclax, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, besimodegib, vorinostat, zanubrutinib, zoledronic acid, amatoxin, anthracycline, anthracene, anthramycin, auristatin, bryostatin, camptothecin, carmofur, combretastatin, cyclosporine, cryptomycin, ecteinascidin, ellipticine, esperamicin, mustine, neothramycin, ozogamicin, phenoxazine, podophyllotoxin, pyrrolobenzodiazepine, sibromycin, tyranstatin, tomatomycin, tubulysin, taxane, vinca alkaloid, 7-epipaclitaxel, 2'-acetylpaclitaxel, 10-deacetylpaclitaxel, 10-deacetyl-7-epipaclitaxel, 7-xylosylpaclitaxel, 10-deacetyl-7-glutaryl paclitaxel, 7-N,N-dimethylglycylpaclitaxel, 7-L-alanyl paclitaxel, lerontaclin, comedocarphin, 9-amino-comedocarphin, 9-nitro-comedocarphin, leptotecan, gimatecan, velotecan, 10-hydroxycamptothecin, 10-hydroxy-7-ethylcamptothecin (SN-38), irinotecan, pimaricin, aclarubicin, sirolimus, tacrolimus, luteinizing hormone, estrogen, rapamycin, pucamycin, trichopilum alkaloid or curcumin.,

[0113] Furthermore, the type of chemotherapy is selected from one or more of amatoxins, anthracyclines, anthracenes, anthramycins, auristatins, bryostatins, camptothecins, carmofur, combretastatins, cyclosporines, cryptomycins, ecteinascidins, ellipticines, esperamicins, maytansines, mustines, neothramycins, ozogamicins, phenoxazines, platinum complexes, podophyllotoxins, pyrrolobenzodiazepines, sibromycins, tyranstatin, tomatomycin, tubulysin, taxanes, or vinca alkaloids.

[0114] In some embodiments, the chemotherapeutic agent is selected from one or more of bleomycin, boanmycin, cabazitaxel, calicheamicin, carboplatin, cisplatin, dactinomycin, daunorubicin, deruxtecan, docetaxel, doxorubicin, epirubicin, eribulin, etoposide, idarubicin, irinotecan, ixabepilone, lurbinectedin, maytansinol, monomethyl auristatin E, monomethyl auristatin F, mitomycin, oxaliplatin, paclitaxel, streptozocin, teniposide, topotecan, trabectedin, valrubicin, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, 7-epipaclitaxel, 2'-acetylpaclitaxel, 10-deacetylpaclitaxel, 10-deacetyl-7-epipaclitaxel, 7-xylosylpaclitaxel, 10-deacetyl-7-glutaryl paclitaxel, 7-N,N-dimethylglycylpaclitaxel, 7-L-alanyl paclitaxel, lerotaxectin, comedocarcin, 9-aminocamptothecin, 9-nitrocamedocarcin, leptotecan, gimatecan, velotecan, 10-hydroxycamptothecin, 10-hydroxy-7-ethylcamptothecin (SN-38), exatecan, vibramycin, aclarubicin, or procarbamptothecin.

[0115] In some embodiments, the ε-poly-L-lysine derivative-drug conjugate, wherein the pharmaceutically active agent residue D is selected from the following structures:

[0116]

Chemical formula

[0117] 。

[0118] In some embodiments, the terminal group X is -OH,

[0119] [Chemical formula]

[0120] or -L 2b -D; said -L 2b -D is preferably

[0121] [Chemical formula] TIFF0007693948000046.tif245169TIFF0007693948000047.tif220169

[0122] .

[0123] In some embodiments, -L 2 -D is selected from any of the following structures:

[0124] [Chemical formula] TIFF0007693948000049.tif246169TIFF0007693948000050.tif228169TIFF0007693948000051.tif225169TIFF0007693948000052.tif245169TIFF0007693948000053.tif240169TIFF0007693948000054.tif231169TIFF0007693948000055.tif245169TIFF0007693948000056.tif228169TIFF0007693948000057.tif245169TIFF0007693948000058.tif233169TIFF0007693948000059.tif232169TIFF0007693948000060.tif245169TIFF0007693948000061.tif245169TIFF0007693948000062.tif238169TIFF0007693948000063.tif245169TIFF0007693948000064.tif245169TIFF0007693948000065.tif245169TIFF0007693948000066.tif234169TIFF0007693948000067.tif246169TIFF0007693948000068.tif233169TIFF0007693948000069.tif245169TIFF0007693948000070.tif220169TIFF0007693948000071.tif245169TIFF0007693948000072.tif220169TIFF0007693948000073.tif226169TIFF0007693948000074.tif245169TIFF0007693948000075.tif222169TIFF0007693948000076.tif236169TIFF0007693948000077.tif222169TIFF0007693948000078.tif246169TIFF0007693948000079.tif233169

[0125] 。

[0126] In some embodiments, -L 2 -D is preferably selected from any of the following structures:

[0127]

Chemical formula

[0128] .

[0129] In some embodiments, the ε-poly-L-lysine derivative-drug conjugate is selected from the following structures:

[0130]

Chemical formula

[0131] .

[0132] In some specific examples, R b is methyl.

[0133] In some specific examples, a = 21 or 22.

[0134] In some specific examples, a = 44 or 45.

[0135] In some specific examples, X is OH.

[0136] In some specific examples, X is L 2b -D.

[0137] In some specific examples, L 2b is a linker containing an enzyme-responsive functional group.

[0138] In some specific examples, L 2b is a linker containing a pH-responsive functional group.

[0139] In some specific examples, D is the residue of a target drug.

[0140] In some specific examples, D is the residue of a chemotherapeutic agent.

[0141] In some specific examples, the ε-poly-L-lysine derivative-drug conjugate is selected from the following structures:

[0142] [Chemical formula] TIFF0007693948000112.tif245169TIFF0007693948000113.tif220169TIFF0007693948000114.tif235169TIFF0007693948000115.tif181169TIFF0007693948000116.tif245169TIFF0007693948000117.tif245169TIFF0007693948000118.tif231169TIFF0007693948000119.tif245169TIFF0007693948000120.tif245169TIFF0007693948000121.tif245169TIFF0007693948000122.tif246169TIFF0007693948000123.tif246169TIFF0007693948000124.tif246169TIFF0007693948000125.tif233169TIFF0007693948000126.tif246169TIFF0007693948000127.tif223169TIFF0007693948000128.tif246169TIFF0007693948000129.tif246169TIFF0007693948000130.tif245169TIFF0007693948000131.tif246169TIFF0007693948000132.tif245169TIFF0007693948000133.tif246169TIFF0007693948000134.tif236169TIFF0007693948000135.tif243169TIFF0007693948000136.tif243169TIFF0007693948000137.tif246169TIFF0007693948000138.tif246169TIFF0007693948000139.tif246169TIFF0007693948000140.tif246169TIFF0007693948000141.tif226169TIFF0007693948000142.tif233169TIFF0007693948000143.tif246169TIFF0007693948000144.tif114169

[0143] 。

[0144] The present invention also provides the following compounds: R 1’ L 2 -D, where R 1 ’ is H, -OH or C1-C3 alkyl, and L 2 and D are as defined above, and the C1-C3 alkyl is preferably methyl.

[0145] The present invention also provides the following compounds:

[0146]

Chemical formula

[0147] 。

[0148] The present invention also provides a compound represented by formula (VI-XV) or a salt thereof:

[0149]

Chemical formula

[0150] Here, P 0 、P 5 、P 6 are each independently selected from OH or a carboxyl protecting group, and P0 , P 5 , P 6 When P is a carboxyl protecting group, they may be the same or different; P 1 , P 2 , P 3 and P 4 are each independently selected from H or an amine protecting group, and when P 1 , P 2 , P 3 and P 4 are amine protecting groups, they are the same or different; In some specific examples, P is a methyl-capped polyethylene glycol residue having a number average molecular weight of 500 to 5,000.

[0151] In some specific examples, L 1 is selected from a covalent bond, C(O) or C(O)-O.

[0152] In some specific examples, L 2a is C(O)-E-C(O), where E is selected from a covalent bond, CH2, CH2-CH2, CH2-CH2-CH2, CH2-O-CH2, CH2-S-CH2, CH2-NH-CH2, CH2-N(Me)-CH2, or CH-CH.

[0153] In some specific examples, n is an integer from 10 to 70.

[0154] In some specific examples, the compound represented by formula (VI-XV) is selected from any of the following compounds:

[0155]

Chemical formula

[0156] .

[0157] In some embodiments, the trifunctional branching center Y is the linker L 0It is linked to an ε-poly-L-lysine residue via [linker], and the molar ratio of the trifunctional branching center Y to the ε-poly-L-lysine structural unit is 0.5:1 to 1.5:1.

[0158] In some embodiments, the trifunctional branching center Y is a linker L 0 It is linked to an ε-poly-L-lysine residue via [linker], and the molar ratio of the trifunctional branching center Y to the ε-poly-L-lysine structural unit is 0.8:1 to 1.2:1.

[0159] In some embodiments, the pharmacokinetic regulator residue P is a linker L 1 It is linked to the trifunctional branching center Y via [linker], and the molar ratio of the pharmacokinetic regulator residue to the ε-poly-L-lysine structural unit is 0.5:1 to 1.5:1.

[0160] In some embodiments, the pharmacokinetic regulator residue P is a linker L 1 It is linked to the trifunctional branching center Y via [linker], and the molar ratio of the pharmacokinetic regulator residue to the ε-poly-L-lysine structural unit is 0.8:1 to 1.2:1.

[0161] In some embodiments, the pharmaceutically active agent residue D is a linker L 2 It is linked to the trifunctional branching center Y via [linker], and the molar ratio of the pharmaceutically active agent residue D to the ε-poly-L-lysine structural unit is 0.5:1 to 1.5:1.

[0162] In some embodiments, the pharmaceutically active agent residue D is a linker L 2 It is linked to the trifunctional branching center Y via [linker], and the molar ratio of the pharmaceutically active agent residue D to the ε-poly-L-lysine structural unit is 0.8:1 to 1.2:1.

[0163] The ε-poly-L-lysine derivative-drug conjugate of the present invention is suitable for the prevention and treatment of diseases such as tumors, inflammation, diabetes, central nervous disorders, cardiovascular disorders, mental diseases, respiratory diseases, ophthalmic diseases, pain, obesity, etc.

[0164] In some embodiments, the ε-poly-L-lysine derivative-drug conjugate is pharmaceutically used for the prevention and treatment of cancer, and the cancer includes breast cancer, ovarian cancer, prostate cancer, melanoma, brain tumor, nasopharyngeal cancer, esophageal cancer, gastric cancer, hepatocellular carcinoma, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, skin cancer, glioblastoma, neuroblastoma, sarcoma, liposarcoma, osteochondrosarcoma, bone cancer, osteosarcoma, testicular tumor, seminoma, uterine tumor, head and neck tumor, multiple myeloma, malignant lymphoma, essential thrombocythemia, leukemia, thyroid tumor, ureteral tumor, bladder tumor, gallbladder cancer, bile duct cancer, choriocarcinoma, or pediatric tumor.

[0165] In some embodiments, the ε-poly-L-lysine derivative-drug conjugate 101B00A (PEG 1K ) DOX of the present invention has an average drug release rate that is 11.6 times that of the dendrimer-drug conjugate 101A1601 (PEG 1K ) (time point: 4 h); the ε-poly-L-lysine derivative-drug conjugate-drug conjugate 101B00B (PEG 2K ) DOX of the present invention has an average drug release rate that is 1.5 times higher than the drug release rate of the prior art dendritic polymer 101A1602 (PEG 2K ) (time point: 4 h). This result shows that compared with the prior art dendritic poly-lysine drug conjugate, the drug conjugate of the present invention with linear ε-poly-L-lysine as the backbone has lower steric hindrance of surface functional groups, promotes the accessibility of enzymes more, has a faster drug release rate, higher regulatability, and promotes the rapid achievement of the effective therapeutic concentration in the initial stage of tumor treatment and the effective suppression of tumor progression more.

[0166] In some embodiments, the average drug release rate of the ε-poly-L-lysine derivative-drug conjugate 101B00E SN-38 of the present invention is 5.6 times higher than that of the dendrimer-drug conjugate 101A1603, which further indicates that the ε-poly-L-lysine derivative-drug conjugate of the present invention has lower steric hindrance and promotes the effective treatment of tumors more.

[0167] In some embodiments, for the tumor / plasma ratio of the drug released from the ε-poly-L-lysine derivative-drug conjugate 101B00E of the present invention, it reached a maximum of 21.2±6.5 in 24 hours, and this result indicated that the ε-poly-L-lysine derivative-drug conjugate of the present invention could be highly concentrated at the tumor site; on the other hand, the tumor / plasma ratio of irinotecan that released the drug showed a maximum value of 4.4±1.2 at 8 hours. Compared with the conventional chemotherapeutic drug irinotecan, the ε-poly-L-lysine derivative-drug conjugate 101B00E of the present invention has better tissue-specific distribution and significant targeted delivery to the tumor site.

[0168] In some embodiments, for the HepG2 hepatocellular carcinoma CDX model, the ε-poly-L-lysine derivative-DOX coupling 101B00A of the present invention showed significantly better tumor inhibition than the dendrimer-DOX coupling 101A1601 and doxorubicin hydrochloride. However, for the dendrimer-DOX coupling 101A1601, when compared with doxorubicin hydrochloride, there was no statistically significant difference in tumor inhibition between the two groups. This result indicated that the ε-poly-L-lysine derivative-DOX conjugate 101B00A with a faster enzyme release rate was more effective in tumor inhibition against the CDX model of HepG2 hepatocellular carcinoma compared with the dendrimer-DOX conjugate 101A1601 with a slower enzyme release rate.

[0169] In some embodiments, for the HepG2 hepatocellular carcinoma CDX model, on the 33rd day, the ε-poly-L-lysine derivative-SN-38 conjugates 101B00C and 101B00E of the present invention showed tumor inhibition rates of 93% and 92% respectively, while the positive drug group, irinotecan, showed a tumor inhibition rate of -49%. From this, it was shown that both the ε-poly-L-lysine derivative-SN-38 conjugates 101B00C and 101B00E have excellent tumor inhibition effects.

[0170] In some embodiments, for the HepG2 hepatocellular carcinoma CDX model, on the 33rd day, the ε-poly-L-lysine derivative-SN-38 conjugates 101C00A and 101D00A of the present invention similarly showed excellent tumor inhibition, showing inhibitions of 90% and 91% respectively. In contrast, irinotecan, a positive drug group, showed a tumor suppression rate of -49%, indicating that the tumor suppression rates of the ε-poly-L-lysine derivative-drug conjugates 101C00A and 101D00A were significantly superior to that of irinotecan, a positive drug.

[0171] In some embodiments, for the BxPC-3 pancreatic cancer subcutaneous transplantation model, 101B00G 5mg / kg (SN-38) was able to achieve the efficacy of paclitaxel albumin 60mg / kg (MTD). By further increasing the amount of 101B00G to 20mg / kg, the inhibitory effect on BxPC-3 pancreatic cancer was further enhanced. Even when 101B00G was administered at 20mg / kg, the impact on the body weight of mice was smaller than that when paclitaxel albumin was administered at 60mg / kg. The effect of 101B00G on BxPC-3 pancreatic cancer was significantly superior to that of paclitaxel albumin, the first-choice drug, suggesting the possibility of clinical application.

[0172] In some embodiments, as a result of tail vein administration of 101B00G 15mg / kg (SN-38) to rats, only during the initial administration period of 0.083 to 2 hours, the ratio of the released SN-38 in plasma to the total SN-38 was relatively high, at 0.16 to 0.03%. After 4 hours to 168 hours of administration (0 hours on the 22nd day corresponds to 168 hours after the third administration), the released SN-38 accounted for 0.012 to 0.015% of the total SN-38, suggesting that 101B00G was extremely stable in rat plasma and did not release SN-38 non-specifically in plasma. And the total SN-38 had a long half-life, with the distribution-phase half-life reaching nearly 4 days, indicating that 101B00G had a long circulation effect in plasma.

[0173] In some embodiments, topotecan administered once a day for 5 days at the MTD dose of the literature, 2 mg / kg (by free base), had a low inhibitory effect on small cell lung cancer of 1,200 mm 3 and the tumor volume on day 22 increased by 22% from the initial value. In contrast, 101B00G continued to reduce the tumor volume even after single administration, and the tumor volume on day 22 decreased by 81% from the initial value. 101B00G was shown to be significantly superior to topotecan, an existing second-line clinical drug for small cell lung cancer, against large tumors of 1,200 mm 3 .

[0174] In some embodiments, for the subcutaneous large tumor model of 600 mm 3 of HepG2 hepatocellular carcinoma, 101B00G continued to reduce the tumor volume after administration, suggesting the potential for clinical application of 101B00G.

[0175] In some embodiments, for the BxPC-3 pancreatic cancer subcutaneous transplantation model, there was a significant difference (T / C = 30%, p = 0.0007) between the nanoalbumin paclitaxel group (60 mg / kg) and the blank control group 21 days after administration, and there was a significant difference (T / C = 7%, p = 0.00006) between 101T00A-O and the blank control group. The efficacy of 101T00A-O was significantly superior to that of the paclitaxel albumin group (60 mg / kg), indicating the potential for clinical application.

[0176] Compared with the prior art, the drug conjugate of the present invention having a linear ε-poly-L-lysine backbone has the following advantages: The polymer drug conjugate provided by the present invention has a longer half-life in vivo compared with the PEG drug conjugate form. When using ε-poly-L-lysine with a degree of polymerization of 25 to 35 as the backbone, 25 to 35 PEG residues can be attached to the conjugate surface, resulting in low renal clearance, low hepatic clearance and splenic clearance, and a long plasma half-life.

[0177] Compared with existing random graft copolymers: Conventional random graft copolymers are troubled by excessive drug coupling and non-uniform drug coupling sites of a single polymer chain. However, the polymer-drug conjugate provided by the present invention has more controllable pharmacokinetic regulation chain segments, drug coupling sites and ratios, and can achieve precise control of the ratio and homogeneity of the drug / pharmacokinetic regulation chain segments. Moreover, by better adjusting and balancing the relationships of drug clearance, drug retention, and drug release, the best therapeutic effect can be achieved.

[0178] Compared with existing dendrimer-drug conjugates, the polymer-drug conjugate provided by the present invention has a small steric resistance of the outermost functional groups, high enzyme accessibility to the linker, and a high drug release rate. Therefore, an effective drug concentration can be accumulated at the lesion site in a short time. Compared with existing dendrimer-drug conjugates, the polymer-drug conjugate provided by the present invention has a better tumor suppression effect on the HepG2 hepatocellular carcinoma CDX tumor model.

[0179] Compared with existing dendrimer drug conjugates, the polymer drug conjugates provided by the present invention are easier to synthesize. The coupling process for obtaining a graft polymer having an average of 60 orthogonally protected functional groups is only carried out once, and appropriate modifications can be continued to exceed the renal clearance threshold and achieve the characteristics of long in vivo circulation. In the synthesis process, the coupling fragment intermediate can achieve a high modification degree of more than 95% without excessive dosing. Since the manufacturing process is short and easy to control, it is suitable for industrial scale-up and manufacturing, and industrial applications can be expected. In addition, the synthetic raw material ε-poly-L-lysine can be mass-produced by microbial fermentation, the production process is mature, and it can be obtained in large quantities. Also, the structure of the raw material ε-poly-L-lysine is clear, and it is easy to establish quality inspection and quality control methods for the starting materials. In contrast, when synthesizing a dendritic poly-L-lysine polymer with 32 lysine residues on the surface, the existing dendritic poly-L-lysine requires 11-step synthetic reactions, and it is necessary to significantly overdose reaction reagents and intermediates. The solvent consumption is large, the synthesis period is long, the cost is high, and it is difficult to establish quality control standards.

[0180] Compared with existing random graft polymer conjugates, the ε-poly-L-lysine backbone drug conjugates of the present invention have controllable group coupling numbers and controllable group coupling sites, and are suitable for coupling drugs for the treatment of various diseases. For example, various target agents, immunosuppressants, immunostimulants, immunomodulators, target proteolytic agents, chemotherapeutic agents, various low-molecular drugs having reactive functional groups, peptide drugs, oligosaccharides, oligonucleotides, lipid drugs, vitamins, enzyme drugs, protein drugs, antibody drugs, etc.

[0181] The terms used in the present invention have the following meanings, unless the opposite is expressed: The term "number of repeating units n" can also be expressed as the degree of polymerization DP. Unless otherwise specified, the number of repeating units n in the present invention represents the number average degree of polymerization, that is, the average value of the number of repeating units contained in the polymer macromolecular chain. When the main chain of ε-poly-L-lysine is synthesized by solid-phase synthesis, the number of repeating units n is a single value. When ε-poly-L-lysine is synthesized by biotransformation or other chemical polymerization, the number of repeating units n has a certain distribution, and its value is represented as a quantitative average value.

[0182] The amine protecting group, hydroxyl protecting group, and carboxyl protecting group of the present invention are groups suitable for amine protection, hydroxyl protection, and carboxyl protection known in the art. Please refer to the literature ("Protective Groups in Organic Synthesis" 5 Th Ed. T. W. Greene & P. G. M. Wuts. P. G. M. Wuts). The term "alkyl" refers to a saturated aliphatic hydrocarbon group containing a straight-chain or branched-chain group of 1 to 30 carbon atoms. Preferably, it is an alkyl group having 1 to 10 carbon atoms, more preferably 1 to 8 carbon atoms. Non-limiting embodiments include, but are not limited to: methyl, ethyl, n-propyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1,1,2-trimethylpropyl, 1-ethyl-2-methylpropyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 3,3-dimethylpentyl, 3,4-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, yukanil, nonyl, decanyl, undecyl, dodecyl, and various isomers thereof.

[0183] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic substituent consisting of 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 6 carbon atoms. Non-limiting embodiments of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, etc., but are not limited thereto. Non-limiting embodiments of polycyclic cycloalkyls include spiro, fused, and bridged cycloalkyls, but are not limited thereto.

[0184] The term "alkenyl" refers to an alkyl as defined in the present invention consisting of at least 2 carbon atoms and at least 1 carbon-carbon double bond, preferably C2-C10 alkenyl, more preferably C2-C6 alkenyl, and examples thereof include vinyl, propenyl, 1-propenyl, etc.

[0185] The term "alkynyl" refers to an alkyl as defined in the present invention consisting of at least 2 carbon atoms and at least 2 carbon-carbon double bonds, preferably C2-C10 alkynyl, more preferably C2-C6 alkynyl, and examples thereof include ethynyl, 1-propynyl, 2-propynyl, etc.

[0186] The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent consisting of 3 to 20 ring atoms, one or more of which are N, O, Si, B, S(O) m 、P(O) ma heteroatom selected from the group consisting of, and m is an integer from 0 to 2. However, the ring moieties of -O-O, -O-S- or -S-S- are excluded, and the remaining ring atoms are carbon. Preferably, it is a monocyclic heteroalkyl group having 3 to 12 ring atoms containing 1 to 4 heteroatoms. Non-limiting examples of the monocyclic heteroalkyl group include pyrrolyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, pyranyl and the like. Examples of the polycyclic heteroalkyl group include spiro ring, macrocycle, and bridged ring heteroalkyl.

[0187] The term "alkoxy" refers to -O-(alkyl) and -O-(cycloalkyl), where alkyl and cycloalkyl are defined as described herein. Non-limiting examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy and the like.

[0188] The term "alkylmercapto" refers to -S-(alkyl) and -S-(cycloalkyl), where alkyl and cycloalkyl are defined as described herein. Non-limiting examples include, but are not limited to, methylmercapto, ethylmercapto, propylmercapto, butylmercapto, cyclopropylmercapto, cyclobutylmercapto, cyclopentylmercapto, cyclohexylmercapto and the like.

[0189] The term "substituted or unsubstituted amino group" refers to NH2, mono-substituted NH2 and bis-substituted NH2, and the mono- or bis-substituent group is preferably, when substituted, deuterium, alkyl, hydroxyl, mercapto, alkenyl, alkynyl, alkoxy, alkanethiol, alkylamino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio group, heterocycloalkylthio, oxo, amino, haloalkyl, hydroxyalkyl, carboxy group or carboxylic acid ester group, etc., independently selected therefrom. The two substituents can be used to form a non-aromatic heterocyclic structure together with the nitrogen atom to which they are attached; The term "aryl" refers to a stable conjugated hydrocarbon ring system group having 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms, which may be a monocyclic, bicyclic, tricyclic or more cyclic aromatic group such as phenyl, naphthyl, anthracene, etc., and the aryl group may include an aryl group on an aryl ring condensed with a heterocycloalkyl group or a cycloalkyl group.

[0190] The term "heteroaryl" refers to an aromatic ring system formed by substituting at least one ring carbon atom with a heteroatom selected from N, O or S, preferably a 5- to 7-membered monocyclic structure or a 7- to 12-membered bicyclic structure, more preferably a 5- to 6-membered heteroaryl group. For example, pyrrolyl, imidazolyl, pyridinyl, pyrimidinyl, thiazolyl, thiophenyl, pyrazinyl, triazolyl, tetrazolyl, oxazolyl, indazolyl, etc. may be mentioned, and the heteroaryl may include a heteroaryl condensed with a heteroaryl, a heterocycloalkyl group or a cycloalkyl ring.

[0191] The term "sulfonyl" is

[0192]

Chemical formula

[0193] refers to, where the substituent is preferably alkyl, alkenyl, alkynyl, amino, alkoxy, alkylmercapto, alkylamino, cycloalkyl, haloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylmercapto or heterocycloalkylmercapto.

[0194] The term "sulfinyl" refers to

[0195]

Chemical formula

[0196] refers to, where the substituent is preferably alkyl, alkenyl, alkynyl, amino, alkoxy, alkylmercapto, alkylamino, cycloalkyl, haloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, or heterocycloalkylmercapto.

[0197] The term "alkylsulfinyl" refers to

[0198]

Chemical formula

[0199] refers to, where the substituent is preferably alkyl as defined above.

[0200] The term "hydroxyl" refers to -OH.

[0201] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0202] The term "nitro" refers to -NO2.

[0203] The term "amino" refers to -NH2.

[0204] The term "cyano" refers to -CN.

[0205] The term "carboxylic acid" refers to -C(O)OH.

[0206] The term "mercapto" refers to -SH.

[0207] The term "carboxylic acid ester" refers to -C(O)O-alkyl, aryl or cycloalkyl, where alkyl, aryl and cycloalkyl are defined as above.

[0208] The term "thioester" refers to -C(O)S-alkyl, aryl or cycloalkyl, where alkyl, aryl and cycloalkyl are defined as above.

[0209] The term "sulfuric acid ester" refers to O-S(O)2O-alkyl, aryl or cycloalkyl, where alkyl, aryl and cycloalkyl are defined as above.

[0210] The term "sulfonic acid ester" refers to S(O)2O-alkyl, aryl or cycloalkyl, where alkyl, aryl and cycloalkyl are defined as above.

[0211] The term "boric acid" refers to B(OH)2.

[0212] The term "borate" refers to B(OR)2, R = alkyl, aryl or cycloalkyl, where alkyl, aryl and cycloalkyl are defined as above. "Substituted" means one or more hydrogen atoms in the group independently substituted with the corresponding number of deuterium or substituents.

[0213] "Pharmaceutically acceptable salt" means a salt that retains the biological effectiveness of the free base without other toxic side effects, which may be an acidic group, a basic group, or an amphoteric group, and non-limiting embodiments include, but are not limited to, the following: Acid salts include hydrochloride, hydrobromide, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, nitrate, acetate, propionate, caprylate, caprate, formate, acrylate, isobutyrate, caproate, heptanoate, oxalate, malonate, succinate, suberate, benzoate, methylbenzoate, phthalate, maleate, methanesulfonate, p-toluenesulfonate, benzenesulfonate, (D,L)-tartrate, citrate, maleate, (D,L)-malate, fumarate, stearate, oleate, cinnamate, laurate, glutamate, aspartate, trifluoromethanesulfonate, mandelate, ascorbate, salicylate. When the compound of the present invention contains an acidic group, its pharmaceutically acceptable salts may also include alkali metal salts (e.g., sodium salt or potassium salt), alkaline earth metal salts (e.g., calcium salt or magnesium salt), and organic alkali salts (e.g., alkyl aromatic amines, amino acids, etc.).

[0214] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein, or their physiologically useful salts or prodrugs, together with other chemical components, as well as other components such as physiologically useful carriers and excipients. The pharmaceutical composition is intended to facilitate administration to a living being and to facilitate absorption of the active ingredient for biological activity.

[0215] The abbreviations for protecting groups, amino acids, and other compounds used in the present invention are based on generally used and recognized abbreviations or refer to the IUPAC-IUBC Commission on Biochemical Nomenclature (Biochem. 1972, 11, 942-944) unless otherwise specified.

[0216] [Brief Description of the Drawings] Figure 1 shows the comparison of the enzyme drug release rates of 101A1601 and 101B00A in Biological Safety Evaluation Example A.

[0217] Figure 2 shows the comparison of the enzyme drug release rates of 101A1602 and 101B00B in Biological Safety Evaluation Example B.

[0218] Figure 3 shows the comparison of the enzyme drug release rates of 101B00E and 101A1603 in Biological Safety Evaluation Example C.

[0219] Figure 4 shows the comparison of the enzyme drug release rates of 101C00A and 101D00A in Biological Safety Evaluation Example D.

[0220] Figure 5 shows the SN-38 distribution in plasma / tumor after intravenous injection of irinotecan (calculated based on SN-38, 10 mg / kg) into HepG2 hepatocellular carcinoma CDX model mice in Biological Safety Evaluation Example E.

[0221] Figure 6 shows the distribution of piperazine-10-O-SN38 (10112) in plasma / tumor after intravenous injection of 101B00E (calculated based on SN38, 10 mg / kg) into HepG2 hepatocellular carcinoma CDX model mice in Biological Safety Evaluation Example E.

[0222] Figure 7 shows the SN-38 ratio (SN-38) in tumor / plasma after intravenous injection of 101B00E and irinotecan 10 mg / kg into HepG2 hepatocellular carcinoma CDX model mice in Biological Safety Evaluation Example E (101B00E was calculated by converting the measured released piperazine-10-O-SN38 (10112) to SN-38).

[0223] Figure 8 shows the inhibitory effects of 101A1601 and 101B00A on the HepG2 hepatocellular carcinoma CDX model in Biological Safety Evaluation Example F.

[0224] Figure 9 shows the inhibitory effects of 101B00C and 101B00E on the HepG2 hepatocellular carcinoma CDX model in Biological Safety Evaluation Example F.

[0225] Figure 10 shows the inhibitory effects of 101C00A and 101D00A on the HepG2 hepatocellular carcinoma CDX model in Biological Safety Evaluation Example F.

[0226] Figure 11 shows the comparison of the effectiveness of 101B00G and nanoalbumin paclitaxel against the subcutaneous pancreatic cancer transplantation model in Biological Safety Evaluation Example G.

[0227] Figure 12 shows the pancreatic cancer subcutaneous transplantation effects of 101B00G and nanoalbumin paclitaxel on the body weight of the HepG2 liver cancer CDX model in Biological Safety Evaluation Example G. Figure 13 shows the plasma total SN-38 and free SN-38 blood concentration-time curves of the first injection of 101B00G in rats in Biological Safety Evaluation Example H.

[0228] Figure 14 shows the plasma total SN-38 and free SN-38 blood concentration-time curves of the fourth injection of 101B00G in rats in Biological Safety Evaluation Example H.

[0229] Figure 15 shows the comparison of the effectiveness of 101B00G and topotecan against the subcutaneous transplantation model of 1200 mm 3 H69 small cell lung cancer in Biological Safety Evaluation Example I. Figure 16 shows the effectiveness of 101B00G against the subcutaneous transplantation model of 600 mm 3 HepG2 hepatocellular carcinoma in Biological Safety Evaluation Example J.

[0230] Figure 17 shows the comparison of the effectiveness of 101T00A-O and albumin paclitaxel against the subcutaneous pancreatic cancer transplantation model in Biological Safety Evaluation Example K.

[0231] Here, the downward arrow in the figure indicates the administration point.

[0232] [Specific Embodiments] The present invention will be further described by way of examples below, but the present invention is not limited to the scope of the described examples. In the following examples, experimental methods for which specific conditions are not indicated are selected according to conventional methods and conditions or the descriptions in product manuals.

[0233] The abbreviations used in the present invention are shown in the following table:

[0234] [Table 1] TIFF0007693948000174.tif221169TIFF0007693948000175.tif247169

[0235] The present invention has been described according to its specific embodiments, but specific modifications and equivalent variations will be apparent to those skilled in the art and are included within the scope of the present invention.

[0236] Analysis method: HPLC analysis method A: Equipment: Agilent 1260 HPLC Column: Ultimate XB-C8 (3μm 300A 100mm), column temperature 40°C Phase A: 0.05% trifluoroacetic acid / water, Phase B: 0.05% trifluoroacetic acid / acetonitrile, flow rate 0.4 ml / min, detection wavelength 210 nm, gradient 0.0 - 10 minutes 95 - 0% Phase A, 10 - 13 minutes 0% Phase A, 13 - 14 minutes 0 - 95% Phase A, 14 - 16 minutes 95% Phase A.

[0237] HPLC analysis method B: Equipment: Agilent 1260 HPLC Column: Ultimate XB-C8 (3μm 300A 100mm), column temperature 40°C Phase A: 0.05% trifluoroacetic acid / water, Phase B: 0.05% trifluoroacetic acid / acetonitrile, flow rate 0.4 ml / min, detection wavelength 210 nm, gradient 0.0 - 10 minutes 90 - 0% Phase A, 10 - 15 minutes 0% Phase A, 15 - 16 minutes 0 - 90% Phase A, 16 - 18 minutes 90% Phase A.

[0238] HPLC analysis method C: Equipment: Agilent 1260 HPLC Column: Ultimate XB-C8 (3μm 300A 100mm), column temperature 40°C Phase A: 0.05% trifluoroacetic acid / water, Phase B: 0.05% trifluoroacetic acid / acetonitrile, flow rate 0.4 ml / min, detection wavelength 210 nm, gradient 0.0 - 10 min 90 - 0% Phase A, 10 - 14 min 0% Phase A, 14 - 15 min 0 - 90% Phase A, 15 - 17 min 90% Phase A.

[0239] HPLC analysis method D: Equipment: Agilent 1260 HPLC Column: Ultimate XB-C18 (4.6μm 300A 250mm), column temperature 40°C Phase A: 0.05% trifluoroacetic acid / water, Phase B: 0.05% trifluoroacetic acid / acetonitrile, flow rate 0.1 ml / min, detection wavelength 210 nm, gradient 0.0 - 20 min 95 - 0% Phase A, 20 - 25 min 0% Phase A, 25 - 25.2 min 0 - 95% Phase A, 25.2 - 28 min 95% Phase A.

[0240] Example 1: Synthesis of Compound 10101

[0241]

Chemical formula

[0242] Dissolve Boc-Lys(Boc)-OH (0.42 kg) and p-nitrophenol (192 g) in EtOAc (12 L) and cool to 0°C in an ice-salt bath. Dissolve DCC (284 g) in EtOAc (6 L) and add dropwise to the above solution. Continue the reaction at 0°C for 2 hours. Warm to room temperature, carry out the reaction overnight, filter, wash the filter cake with cold EtOAc, and concentrate the filtrate. Wash the product with petroleum ether / ether, dissolve it in EtOAc, and then precipitate it again with petroleum ether / ether. Filter and spin-dry to obtain Boc-Lys(Boc)-pNB(10101) as a white powder.

[0243] 1 H NMR (400 MHz, DMSO-d6) δ 8.37 ‐ 8.28 (m, 2H), 7.56 (d, J = 6.8 Hz, 1H), 7.44 ‐ 7.35 (m, 2H), 6.82 (t, J = 5.7 Hz, 1H), 4.16 (dt, J = 12.0, 6.2 Hz, 1H), 2.93 (q, J = 6.1 Hz, 2H), 1.90 ‐ 1.66 (m, 2H), 1.39 (d, J = 14.9 Hz, 22H). Example 2: Synthesis of Compound 10102

[0244]

Chem.

[0245] Boc-Lys(Cbz)-OH (76 g) and p-nitrophenol (32 g) were dissolved in EtOAc (2 L), and the solution was cooled to 2 °C in an ice bath; DCC (47.4 g) was dissolved in EtOAc (1 L) and added dropwise to the above solution, and the reaction was continued at 2 - 3 °C for 2 hours. The reaction mixture was warmed to room temperature, stirred overnight, filtered, and the filtrate was spin-dried. Petroleum ether / methyl tert-butyl ether was added to precipitate the product, which was dried under vacuum to obtain the product Boc-Lys(Cbz)-pNB (10102) as a white powder.

[0246] 1 H NMR (400 MHz, DMSO-d6) δ 8.41 ‐ 8.24 (m, 2H), 7.59 (d, J = 6.8 Hz, 1H), 7.44 ‐ 7.25 (m, 8H), 5.01 (s, 2H), 4.23 ‐ 4.06 (m, 1H), 3.02 (q, J = 6.1 Hz, 2H), 1.91 ‐ 1.68 (m, 2H), 1.41 (s, 13H). Example 3: Synthesis of Compound 10103-1k

[0247]

Chem.

[0248] mPEG 1k -OH (25.0 g), KBr (8.06 g), and Na2CO3 (18.55 g) were added to a 1 L flask containing 300 mL of water, and TEMPO (0.78 g) was added. Ice was added until the water temperature dropped to 0 °C. The NaClO solution (74.4 g) was slowly added and reacted for 2 hours. Sodium bisulfite was added to quench the reaction, and it was extracted with dichloromethane, stirred with anhydrous sodium sulfate, and dried for 1 hour. It was filtered, spin-dried, and dried under vacuum to obtain the product mPEG 1k -COOH (10103-1K) 24.00 g, with a yield of 94.6% was obtained.

[0249] 1 H NMR (400 MHz, CDCl3) δ 4.15 (s, 2H), 3.65 (br, 85H), 3.38 (s, 3H). Example 4: Synthesis of Compound 10103-2k

[0250]

Chemical Structure

[0251] mPEG 2k -OH (50.0 g), KBr (2.083 g), and Na2CO3 (18.55 g) were added to a 1 L flask containing 300 mL of water, and TEMPO (0.78 g) was added. Ice was added for the ice bath, and when the water temperature dropped to 0 °C, the NaClO solution (74.4 g) was slowly added. It was reacted for 2 hours, sodium bisulfite was added to quench the reaction, extracted with dichloromethane, washed with saturated NaCl, and the aqueous phase was clarified. Dried with anhydrous sodium sulfate, filtered, and the filtrate was spin-dried and dried under vacuum to obtain the white solid mPEG 2k -COOH (10103-2K) 48.81 g, with a yield of 96.9% was obtained.

[0252] 11H NMR (400 MHz, CDCl3) δ 4.15 (s, 2H), 3.65 (br, 175H), 3.38 (s, 3H). Example 5: Synthesis of Compound 10104-1k

[0253]

Chemical Structure

[0254] 10103A (24.00 g) and NHS (5.44 g) were dissolved in dichloromethane. The solution was cooled to 0 °C in an ice bath. EDCI (9.06 g) was added and the mixture was stirred at room temperature until the precipitate dissolved and the solution became colorless and transparent. The reaction was carried out overnight at room temperature. 120 mL of dichloromethane was added for dilution, and the solution was washed with saturated NaCl, dried over anhydrous sodium sulfate, spin-distilled, and then dried under vacuum. The product mPEG 1k -NHS (10104-1K) 23.46 g was obtained with a yield of 89.2%.

[0255] 1 1H NMR (400 MHz, CDCl3) δ 4.53 (s, 2H), 3.64 (br, 86H), 3.38 (s, 3H), 2.86 (s, 4H). Example 6: Synthesis of Compound 10104-2k

[0256]

Chemical Structure

[0257] 10103 (48.81 g) and NHS (5.54 g) were dissolved in dichloromethane and cooled to 0 °C in an ice bath. EDCI (9.23 g) was added and the mixture was stirred. The reaction was carried out overnight at room temperature. 250 mL of dichloromethane was added for dilution, and the solution was washed with water and saturated NaCl, dried over anhydrous sodium sulfate, spin-dried, and then dried under vacuum to obtain the product mPEG 2k -NHS (10104-2K) 48.34 g was obtained with a yield of 94.5%.

[0258] 11H NMR (400 MHz, CDCl3) δ 4.53 (s, 2H), 3.65 (br, 178H), 3.39 (s, 3H), 2.87 (s, 4H). Example 7: Synthesis of Compound 10105

[0259]

Chemical Structure

[0260] Fmoc-Val-OH (10.17 g) and NHS (3.89 g) were dissolved in dichloromethane (200 ml), stirred in an ice bath for 15 minutes, DCC (6.59 g) was added portionwise several times, and the mixture was stirred at room temperature for about 5 hours. After filtration, the filtrate was spin distilled to remove the solvent, and a transparent viscous substance was obtained, which was dissolved by adding DME (80 ml). 101M07 (5.78 g) was dissolved in water (80 ml), NaHCO3 (2.78 g) was added, and THF (44 ml) was added. The resulting solution was added to the DME solution of the above product, and the reaction was carried out for 12 hours with vigorous stirring. The pH was adjusted to about 4 with 2N hydrochloric acid, a large amount of white precipitate was deposited, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. The product Fmoc-Val-Cit-OH (10105) (10.15 g) was obtained by spin drying. LCMS (m / z): [M+H] + = 496.9。

[0261] Example 8: Synthesis of Compound 10106

[0262]

Chemical Structure

[0263] Fmoc-Val-Cit-OH (9.93 g) was dissolved in dichloromethane / MeOH (200 / 100 ml), PABOH (4.93 g) was added, EEDQ (4.9 g) was added, protected with nitrogen, and stirred at 30 °C. After 1 hour of reaction, EEDQ (5 g) was continuously added, and the reaction was continued for 12 hours. Spin distillation was performed to remove most of the solvent, 90 ml of ethyl acetate was added, filtered, and the product Fmoc-Val-Cit-PABC(10106) 8.80 g (14.6 mmol, 73.1%) was obtained by vacuum drying. LCMS (m / z): [M+H] + = 601.9.

[0264] Example 9: Synthesis of Compound 10107

[0265]

Chemical Structure

[0266] Fmoc-Val-Cit-PABC (8.80 g) was dissolved in DMF (170 ml), ice-bathed, then bis(p-nitrophenyl) carbonate (13.32 g) was added, DIPEA (5.31 ml) was added dropwise, and the mixture was warmed to room temperature while stirring for 1 hour. DMF was removed by spin distillation, dichloromethane / EtOAc was added, filtered, and Fmoc-Val-Cit-PABC-pNP(10107) 9.40 g (12.1 mmol, 82.9%) was obtained as a pale yellow solid by vacuum drying.

[0267] LCMS (m / z): [M+H] + = 766.9 Example 10: Synthesis of Compound 10108

[0268]

Chemical Structure

[0269] Fmoc-Val-Cit-PABC (1.23 g, 1 eq.) was dissolved in DMF (13 mL), DOX-HCl (0.93 g) was added, DIPEA (0.836 mL) was added, and the reaction was carried out at room temperature for 2 hours. 120 mL of MTBE was added to obtain a reddish-brown precipitate, which was filtered. The filter cake was washed with MTBE / MeOH and dried in vacuo to obtain 1.85 g (98.9%) of the product Fmoc-Val-Cit-PABC-DOX (10108).

[0270] LCMS: Theoretical [M+2H] 2+ = 586.2, Found 584.2.

[0271] Example 11: Synthesis of Compound 10109

[0272]

Chemical Structure

[0273] Fmoc-Val-Cit-PABC-DOX (1.85 g) was dissolved in DMF (18 mL), and diethylamine (2 mL) was added. The solution turned purple-black, stirred at room temperature for 15 minutes, 280 mL of MTBE was added, filtered, and 1.47 g of dark red solid powder NH2-Val-Cit-PABC-DOX (10109) was obtained by drying in vacuo. It was prepared by prep-HPLC and freeze-dried to obtain 1.12 g of bright red powder (74%).

[0274] 11H NMR (400 MHz, DMSO-d6) δ 14.03 (s, 1H), 13.27 (s, 1H), 10.17 (s, 1H), 8.67 (d, J = 7.6 Hz, 1H), 8.09 (d, J = 5.1 Hz, 3H), 7.91 (d, J = 4.9 Hz, 2H), 7.65 (p, J = 3.8 Hz, 1H), 7.54 (d, J = 8.1 Hz, 2H), 7.26 (d, J = 8.2 Hz, 2H), 6.83 (d, J = 8.1 Hz, 1H), 6.07 (t, J = 5.9 Hz, 1H), 5.47 (d, J = 7.9 Hz, 3H), 5.22 (d, J = 3.6 Hz, 1H), 4.92 (d, J = 17.6 Hz, 4H), 4.72 (s, 1H), 4.58 (s, 2H), 4.49 (td, J = 8.2, 5.1 Hz, 1H), 4.16 (q, J = 6.6 Hz, 1H), 3.99 (s, 3H), 3.80 ‐ 3.60 (m, 2H), 3.11 ‐ 2.87 (m, 5H), 2.26 ‐ 1.97 (m, 4H), 1.84 (td, J = 13.0, 3.9 Hz, 1H), 1.71 (dq, J = 13.3, 7.2, 6.5 Hz, 1H), 1.61 (dtd, J = 13.5, 8.7, 4.6 Hz, 1H), 1.52 ‐ 1.34 (m, 3H), 1.13 (d, J = 6.4 Hz, 3H), 0.93 (dd, J = 6.8, 2.4 Hz, 7H). LC-MS: [M+H] + =949.2 Example 12: Synthesis of Compound 10126

[0275]

Chem.

[0276] SN-38 (2.00 g) was dispersed in 80 mL of DMF, 3.18 mL of DIPEA was added to form a yellow suspension, TBSCl (2.69 g) was added, and the solution gradually became clear. The reaction was carried out at room temperature for about 1 hour. 80 mL of water was added dropwise under an ice bath to precipitate the product, which was filtered. The filter cake was washed twice with water and dried under vacuum at 40 °C overnight. 10-O-TBS-SN-38 (10126), 2.42 g (93.9%) was obtained as a pale yellow solid. LCMS: [M+H] + =507.2 Example 13: Synthesis of Compound 10127

[0277]

Chemical Structure

[0278] 10-O-TBDMS-SN-38 (173 mg) and DMAP (136 mg) were suspended in dichloromethane (2 ml), and a solution of dichloromethane (3.5 ml) containing triphosgene (53 mg) was added. After the solution became turbid, the reaction was carried out at 20 °C for 10 minutes. Fmoc-Val-Cit-PABC (180 mg) was added. After the solution became clear, the reaction was continued. The reaction was carried out overnight at room temperature. By column chromatography with 5-20% MeOH / dichloromethane, the product 20-O-(Fmoc-Val-Cit-PABC)-10-O-TBS-SN-38 (10127) 264 mg (46.5%) was obtained.

[0279] LCMS: Theoretical [M+2H] 2+ =567.7, Measured 565.2.

[0280] Example 14: Synthesis of Compound 10128

[0281]

Chemical Structure

[0282] 20-O-(Fmoc-Val-Cit-PABC)-10-O-TBS-SN-38(10127) (200 mg) was dissolved in DMF (6 ml), piperidine (300 μl) was added, and the mixture was reacted at room temperature for 20 minutes. The product was precipitated by the addition of methyl tert-butyl ether and dried under vacuum to obtain 140 mg (99.0%) of the product NH2-Val-Cit-PABC-20-O-SN-38(10128).

[0283] LCMS: Theoretical [M+H] + =798.1 Comparative Example 1: Synthesis of Compound 101A1601 Synthesis of Compound 101A01

[0284]

Chemical Structure

[0285] To a CH3CN / DMF solution (20 / 10 mL) of diphenylmethylamine (BHA, 2.50 g), DIPEA (2.53 g) was added, and then 10101 (7.02 g) was added portionwise several times under a nitrogen atmosphere in a 5 °C ice bath. The reaction solution was stirred for 12 hours. 0.5 N aqueous sodium hydroxide solution (100 mL) was added to the above reaction solution, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and 3.8 g of the crude product BHA-Lys(α,ε-Boc)(101A01) was obtained with a yield of 55%. MS (ESI), m / z, 512.2 [M+H] + . Synthesis of Compound 101A02:

[0286]

Chemical Structure

[0287] To a dichloromethane solution (10 mL) of 101A01 (3.80 g) was added TFA (2 mL), and the mixture was stirred for 12 hours at room temperature (25 °C) under nitrogen protection. MTBE (30 mL) was added to the above reaction solution for concentration, and dichloromethane (10 mL) dissolved by addition was added and concentrated to obtain the product 101A02, BHA-Lys(α,ε-NH3+CF3COO-) (101A02), as a foamy solid, 3.0 g, in 89% yield. MS (ESI), m / z, 312.1 [M+H] + 。

[0288] Synthesis of compound 101A03:

[0289]

Chemical formula

[0290] To a DMF solution (10 mL) of 101A02 (TFA salt, 1.00 g) was added DIPEA (1.70 g), and then 10101 (1.91 g) was added portionwise under a 5 °C ice-water bath and nitrogen protection. The reaction solution was stirred for 12 hours. The above reaction solution was poured into a 0.5 N aqueous sodium hydroxide solution (50 mL), stirred for 1 hour, filtered, and the filter cake was washed with a 0.5 N aqueous sodium hydroxide solution. The filter cake was dried in vacuo to obtain the product 101A03 as a white solid, 1.50 g, in 84% yield. MS (ESI), m / z, 968.5 [M+H] + 。

[0291] Synthesis of compound 101A04:

[0292]

Chemical formula

[0293] To a dichloromethane solution (10 mL) of 101A03 (1.5 g, 1.55 mmol) was added TFA (2 mL), and the mixture was stirred for 12 hours at room temperature (25 °C) under nitrogen protection. The above reaction solution was concentrated, and the product 101A04 was obtained as a foamy solid, 1.50 g, in 95% yield with dichloromethane.

[0294] 1 1H NMR (400 MHz, D2O) δ 7.41 ‐ 7.19 (m, 10H), 6.03 (s, 1H), 4.36 (t, J = 7.4 Hz, 1H), 3.94 (t, J = 6.5 Hz, 1H), 3.85 ‐ 3.76 (m, 1H), 3.10 (t, J = 7.3 Hz, 2H), 2.90 (t, J = 7.9 Hz, 2H), 2.63 (t, J = 7.7 Hz, 2H), 2.03 ‐ 0.99 (m, 18H). MS (ESI), m / z, 568.3 [M+H]+。

[0295] Synthesis of Compound 101A05:

[0296]

Chem.

[0297] To a DMF solution (30 mL) of 101A04 (TFA salt, 1.50 g), DIPEA (2.60 g) and then 10101 (prepared in Example 1) (3.01 g) were added under a 5 °C ice-water bath and nitrogen protection, and the reaction solution was stirred and reacted for 12 hours. The above reaction solution was poured into a 0.5 N aqueous sodium hydroxide solution (100 mL) and stirred for 1 hour. The product 101A05 was filtered, the solid was collected, and by vacuum drying, 2.3 g of the product 101A05 as a white solid was obtained in a yield of 83%.

[0298] MS (ESI), m / z, 941.0 [M / 2+H]+。

[0299] Synthesis of Compound 101A06:

[0300]

Chem.

[0301] To a dichloromethane solution (10 mL) of 101A05 (2.30 g) was added TFA (10 mL), and the mixture was stirred at room temperature (25 °C) under nitrogen protection for 4 hours. The above reaction solution was concentrated, and dichloromethane was dissolved in dichloromethane / MeOH with MTBE added for concentration to obtain product 101A06 as a foamy solid, 2.41 g, in a yield of 98.5%. MS (ESI), m / z, 540.8 [M / 2+H] + 。

[0302] 1 H NMR (400 MHz, D2O) δ 7.45 ‐ 7.11 (m, 10H), 6.02 (s, 1H), 4.32 ‐ 4.09 (m, 3H), 4.01 ‐ 3.92 (m, 2H), 3.85 (t, J = 6.7 Hz, 1H), 3.78 (t, J = 6.7 Hz, 1H), 3.22 ‐ 2.80 (m, 14H), 1.94 ‐ 1.06 (m, 42H). Synthesis of compound 101A07:

[0303]

Chemical formula

[0304] To a DMF solution (20 mL) of 101A06 (TFA salt, 2.40 g) were added DIPEA (3.73 g), and then 10101 (prepared in Example 1) (5.41 g) was added under a 5 °C ice-water bath and nitrogen protection, and the reaction solution was stirred and reacted for 12 hours. The above reaction solution was poured into a 0.5 N aqueous sodium hydroxide solution (150 mL) and stirred for 1 hour. It was filtered, and the solid dissolved in acetonitrile was recovered, water (1 / 1 volume) was added to precipitate, stirred for 1 hour, filtered, the solid was recovered, and vacuum dried to obtain product 101A07 as a white solid, 3.82 g, in a yield of 85%.

[0305] Synthesis of compound 101A08:

[0306]

Chemical formula

[0307] TFA (10 mL) was added to a dichloromethane solution (10 mL) of 101A07 (3.0 g), and the mixture was stirred at room temperature (25° C.) under nitrogen protection for 4 hours. The above reaction solution was concentrated, dissolved in acetonitrile (30 mL), and precipitated by adding MTBE (150 mL), filtered, and the solid was collected and dried in vacuum to obtain product 101A08, 2.9 g, 91% yield, as a white solid. MS (ESI), m / z, 702.7 [M / 3+H] + .

[0308] 1 H NMR (400 MHz, D2O) δ 7.64 - 6.97 (m, 10H), 6.02 (s, 1H), 4.31 - 4.04 (m, 7H), 3.96 (t, J = 6.7 Hz, 4H), 3.90 - 3.79 (m, 5H), 3.30 - 2.77 (m, 30H), 1.93 - 0.96 (m, 90H). Synthesis of compound 101A09:

[0309] [ka]

[0310] DIPEA (3.94 g) was added to a DMF solution (20 mL) of 101A08 (TFA salt, 2.50 g), and then 10101 (5.94 g) was added under the protection of nitrogen and ice-water bath at 5°C, and the reaction solution was stirred for 12 hours. The above reaction solution was poured into 0.5N aqueous sodium hydroxide solution (100 mL), stirred for 1 hour, filtered, and the solid was collected and dried in vacuum to obtain product 101A09, 3.7 g as a white solid, 80% yield.

[0311] Synthesis of compound 101A10:

[0312] [ka]

[0313] TFA (10 mL) was added to a dichloromethane solution (10 mL) of 101A09 (3.0 g), and the mixture was stirred at room temperature (25 °C) under nitrogen protection for 12 hours. The above reaction solution was concentrated, dissolved in acetonitrile (20 mL), MTBE (100 mL) was added to cause precipitation, and the mixture was filtered. The solid was recovered and dried under vacuum to obtain 3.0 g of product 101A10 as a white solid, with a yield of 94%. MS (ESI), m / z, 832.1 [M / 5+H] + 。

[0314] 1 H NMR (400 MHz, D2O) δ 7.45 ‐ 7.03 (m, 10H), 6.01 (s, 1H), 4.34 ‐ 4.06 (m, 16H), 4.02 ‐ 3.92 (m, 8H), 3.90 ‐ 3.79 (m, 8H), 3.29 ‐ 2.72 (m, 62H), 2.00 ‐ 0.93 (m, 186H). Synthesis of compound 101A11:

[0315]

Chemical formula

[0316] DIPEA (3.3 g) was added to a DMF solution (20 mL) of 101A10 (TFA salt, 2.0 g). Then, 10102 (4.79 g) was added under a 5 °C ice-water bath and nitrogen protection, and the reaction solution was stirred and reacted for 6 hours. The above reaction solution was poured into a 0.5 N aqueous sodium hydroxide solution (100 mL), stirred for 1 hour, filtered, and the solid was recovered. An aqueous solution (200 mL) was added, stirred for 1 hour, and filtered. This was repeated twice, the solid was recovered, and dried under vacuum to obtain 3.8 g of product 101A11 as a white solid, with a yield of 94%.

[0317] Synthesis of compound 101A12:

[0318]

Chemical formula

[0319] To a solution of 101A11 (2.50 g, 0.16 mmol) in acetic acid (50 mL, if not clarified, diatomaceous earth filtration is required), Pd / C or Pd(OH)2 (10%, 1.0 g, 10 mL) was added at room temperature (25 °C), protected with nitrogen, replaced with hydrogen gas to form a hydrogen atmosphere (hydrogen balloon), and stirred for 24 hours (30 °C). The above reaction solution was concentrated, dissolved in methanol (20 mL), MTBE (100 mL - 200 mL) was added to precipitate, filtered, the solid was recovered, and by vacuum drying, 1.7 g of product 101A12, as an off-white solid, was obtained with a yield of 80%. MS (ESI), m / z, 832.1 [M / 5 + H] + 。

[0320] 1 H NMR (400 MHz, D2O) δ 7.53 - 6.98 (m, 10H), 6.02 (s, 1H), 4.15 (s, 34H), 3.89 (d, J = 26.3 Hz, 38H), 3.38 - 2.71 (m, 126H), 1.86 (s, 118H), 2.21 - 0.70 (m, 669H). Synthesis of Compound 101A13 - 1K:

[0321]

Chemical Structure

[0322] To a solution of 101A12 (1.0 g) in DMF (10 mL), DIPEA (0.96 g) was added, followed by 10104 - 1K (3.6 g). The reaction solution was stirred and reacted for 12 hours at 25 °C under nitrogen protection. MTBE / MeOH was added, filtered, and the filter cake was vacuum dried to obtain the crude product 101A13 - 1K, which was directly used in the next step of the reaction.

[0323] Synthesis of Compound 101A14 - 1K:

[0324]

Chemical Structure

[0325] TFA (20 mL) was added to a dichloromethane solution (30 mL) of 101A13-1K, and the mixture was stirred and reacted at room temperature (25 °C) under nitrogen protection for 12 hours. The reaction solution was concentrated, MTBE (50 mL) was added, and the mixture was stirred for 10 minutes, filtered, and the filter cake was dried under vacuum to obtain 2.5 g of the product 101A14-1K as a colorless oil in a yield of 72% in two steps.

[0326] Synthesis of compound 101A15-DGA-1K:

[0327]

Chemical formula

[0328] DIPEA (277 mg) and then diethylene glycol anhydride (159 mg) were added to a DMF solution (5 mL) of 101A14-1K, and the reaction solution was stirred and reacted at 25 °C under nitrogen protection for 12 hours. MTBE was added and the mixture was stirred for 10 minutes, the supernatant was decanted, and the residue was dried under vacuum to obtain 0.8 g of the product 101A15-DGA-1K as an off-white solid in a yield of 74%.

[0329] Synthesis of compound 101A1601:

[0330]

Chemical formula

[0331] To a DMF solution (5 mL) of 101A15-DGA-1K (300 mg, 0.006 mmol), DIPEA (97 mg, 0.76 mmol, 128.0 eq), 10109 (prepared in Example 11) (269 mg, 0.28 mmol, 48.0 eq) and PyBOP (246 mg, 0.47 mmol, 80.0 eq) were added under a 5°C ice bath and nitrogen protection, and the solution was stirred and reacted for 12 hours. The complete reaction was monitored by HPLC, and the residue was directly dissolved in DMSO / MeOH, purified by ultrafiltration, and freeze-dried to obtain 300 mg of the final product (101A1601), a dark purple solid, with a purity of 98% or more.

[0332] HPLC purity measurement method: Dissolve the compound in methanol; Instrument: Agilent 1260 HPLC; Column: Ultimate XB-C8 (3 μm 300A 100 mm); Column temperature 40°C; Flow rate 0.4 mL / min; Detection wavelength UV 210 nm; Area calculation method; Mobile phase A: 0.05% TFA in water; Mobile phase B: 0.05% TFA in CH3CN; Gradient: For 16 minutes, for 10 minutes: 95:5 A / B ~ 0:100 A / B, for 5 minutes: 0:100 A / B, for 1 minute: 0:100 A / B ~ 95:5 A / B.

[0333] Comparative Example 2: Synthesis of compound 101A1602: Synthesis of compound 101A13-2K

[0334]

Chemical formula

[0335] To a DMF solution (10 mL) of 101A12 (1.0 g), DIPEA (0.96 g) was added, followed by 10104-2K (6.48 g). Under nitrogen protection at 25°C, the reaction solution was stirred and reacted for 12 hours. MTBE (100 mL) was added to the above reaction solution, filtered, and the cake was vacuum dried to obtain 4.0 g of the product 101A13-2K, with a yield of 70%.

[0336] Synthesis of compound 101A14-2K

[0337]

Chem.

[0338] TFA (30 mL) was added to a dichloromethane solution (50 mL) of 101A13 - 2K (4.0 g), and the mixture was stirred at room temperature (25 °C) under nitrogen protection for 12 hours. MTBE was added, and the mixture was cooled to room temperature - 20 °C to precipitate crystals, which were filtered. The solid was recovered and dried under vacuum to obtain 3.8 g of the product 101A14 - 2K as an off - white solid, with a yield of 95%.

[0339] Synthesis of Compound 101A15 - DGA - 2K

[0340]

Chem.

[0341] To a DMF solution (5 mL) of 101A14 - 2K (0.8 g), DIPEA (133 mg) was added, followed by diethylene glycol anhydride (76 mg). The reaction solution was stirred and reacted at 25 °C under nitrogen protection for 12 hours. CH3CN / MTBE was added and stirred for 10 minutes, and the supernatant was decanted. The residue was dried under vacuum to obtain 0.7 g of the product 101A15 - DGA - 2K as an off - white solid, with a yield of 83%.

[0342] Synthesis of Compound 101A1602:

[0343]

Chem.

[0344] To a DMF solution (5 mL) of 101A15-DGA-2K (500 mg, 0.006 mmol), DIPEA (101 mg, 0.78 mmol, 128.0 eq), 10109 (prepared in Example 11) (278 mg, 0.29 mmol, 48.0 eq), and PyBOP (254 mg, 0.49 mmol, 80.0 eq) were added under a 5°C ice-water bath and nitrogen protection, and the solution was stirred and reacted for 12 hours. The complete reaction was monitored by HPLC, MTBE (50 mL) was added, stirred for 10 minutes, the supernatant was decanted, the residue was directly dissolved in DMSO / MeOH, purified by ultrafiltration, and freeze-dried to obtain 500 mg of the final product (101A1602) as a dark purple solid with a purity of 98% or higher.

[0345] HPLC method for purity measurement: Dissolve the compound in methanol; Instrument: Agilent 1260 HPLC; Column: Ultimate XB-C8 (3 μm 300A 100 mm); Column temperature 40°C; Flow rate 0.4 mL / min; Detection wavelength UV 210 nm; Area calculation method; Mobile phase A: 0.05% TFA in water; Mobile phase B: 0.05% TFA in CH3CN; Gradient: 16 minutes, 0 - 10 minutes: 5% B - 100% B, 10 - 15 minutes: 100% B, 15 - 15.1 minutes 100% B - 5% B, 15.1 - 16 minutes 5% B.

[0346] Comparative Example 3: Synthesis and property evaluation of compound 101A1603 Synthesis of compound 101A15-SA-2K

[0347]

Chemical formula

[0348] To a DMF solution (5 mL) of 101A14-2K (1.0 g, 0.013 mmol), DIPEA (166 mg, 1.29 mmol, 100.0 eq) was added, followed by succinic anhydride (82 mg, 0.82 mmol, 64.0 eq). The reaction solution was stirred and reacted at 25 °C under nitrogen protection for 12 hours. After the reaction was completed, MTBE (50 mL) was added and stirred for 10 minutes. The supernatant was decanted and repeated 3 times. The residue was dried under vacuum to obtain the product 101A15-SA-2K, 0.8 g, with a yield of 76%.

[0349] Synthesis of Compound 101A1603

[0350]

Chemical Structure

[0351] To a DMF solution (5 mL) of 101A15-SA-2K (500 mg, 0.006 mmol), DIPEA (101 mg, 0.78 mmol, 128.0 eq), 10114 (269 mg, 0.30 mmol, 48.0 eq) and PyBOP (256 mg, 0.49 mmol, 80.0 eq) were added under a 5 °C ice bath and nitrogen protection. The solution was stirred and reacted at room temperature for 12 hours. The complete reaction was monitored by HPLC. MTBE (50 mL) was added and stirred for 10 minutes. The upper clear liquid was decanted. The residue was directly dissolved in DMSO / MeOH, purified by ultrafiltration, and freeze-dried to obtain 592 mg of the final product (101A1603), as a pale yellow solid, with an HPLC purity of over 90%.

[0352] Measurement of SN-38 Coupling of Compound 101A1603 Papain was weighed and added to PBS to form a PBS solution of 6.25 mg / ml.

[0353] Weighed 122.8 mg of freeze-dried BHA-Lys32(α-SA-VC-PABC-piperazine-10-O-SN-38, ε-mPEG2k)32(101A1603), added 4.910 ml of deionized water to obtain a polymer solution of 25.0 mg / ml.

[0354] Took 0.200 ml of the polymer solution + 1.800 ml of papain PBS solution, the final concentration of the polymer was 2.50 mg / ml, and it was shaken in a 37 °C water bath for 72 hours. Cooled to room temperature, pipetted 0.100 ml, added 0.900 ml of DMSO, characterized the peak area of piperazine-10-O-SN-38 after enzymatic digestion by HPLC, and calculated the concentration of TFA-piperazine-10-O-SN-38 using the standard curve of TFA-piperazine-10-O-SN-38, which was 552.6 μg / ml. After conversion, the concentration of SN-38 after enzymatic hydrolysis of 5.00 mg / ml polymer was 267.1 μg / ml, and the coupling amount of SN-38 in BHA-Lys32(α-SA-VC-PABC-piperazine-10-O-SN-38, ε-mPEG2k)32 was calculated to be 10.7 wt.%.

[0355] Example 15: Synthesis of Compound 101B00A Synthesis of Compound 101B01

[0356]

Chemical Structure

[0357] Suspended ε-poly-L-lysine hydrochloride (4.792 g, 29.09 mmol, number of moles of structural unit) in 100 g of DMSO, added triethylamine (8.89 g, 87.27 mmol), added α-Boc-ε-Cbz-L-lysine-NHS activated ester (10102, prepared in Example 2) (20.83 g, 46.63 mmol), stirred at 30 °C for 13 hours under nitrogen protection to complete the reaction. Put the reaction solution into a beaker, added 800 ml of ACN, filtered, washed the filter cake with acetonitrile, water, and acetonitrile in sequence, and dried in vacuo to obtain the product ε-PolyLys30 -[Lys(α-Boc, ε-Cbz)]30(101B01) was obtained as a white solid (12.13 g, 85%).

[0358] 1 H NMR (400 MHz, DMSO-d6) 7.74 (m, 62H), 7.27 (m, 179H), 6.90 (m, 28H), 5.16 ‐ 4.80 (s, 60H), 4.31 ‐ 4.01 (br, 30H), 4.00 ‐ 3.68 (br, 30H), 3.12 ‐ 2.82 (m, 120H), 1.91 ‐ 0.58 (m, 642H). The experiment was repeated three times to obtain three batches with a constant amount of side-chain coupling.

[0359]

Table 2

[0360]

Table 3

[0361] Synthesis of Compound 101B02

[0362]

Chemical formula

[0363] α-[Boc-Lys(Cbz)] 30 -ε-PolyLys 30 (101B01) (2.92 g) was dissolved in 30 ml of acetic acid + 30 ml of methanol, heated to dissolve, 591 mg (10%) of palladium carbon was added together with hydrogen, and stirred at 25 °C for 18 hours. Celite was added, filtered, the filtrate was concentrated by spin distillation, precipitated with methyl tert-butyl ether, and by vacuum drying, α-[Boc-Lys(NH3 + COO - )] 30 -ε-PolyLys 30(101B02) was obtained as a white powder.

[0364] 1 H NMR (400 MHz, Deuterium Oxide) δ 4.20 - 3.98 (m, 30H), 3.88 (t, J = 7.0 Hz, 31H), 3.04 (s, 61H), 2.87 (t, J = 7.6 Hz, 59H), 1.84 (s, 122H), 1.78 - 1.10 (m, 594H). Synthesis of Compound 101B03

[0365]

Chemical Structure

[0366] 101B02 (2.00 g) was dissolved in 9.12 ml of water, 10104-1K (3.42 g) was dissolved in 13.7 ml of acetonitrile, the two solutions were mixed, 1.19 ml of DIPEA was added, stirred overnight at room temperature, the solvent was removed by spin distillation, extracted with dichloromethane, dried over anhydrous sodium sulfate, and spin-dried to obtain 8.69 g of product 101B03.

[0367] 1 H NMR (400 MHz, Deuterium Oxide) δ 4.25 (br, 30H), 4.09 (br, 96H), 3.93 - 3.45 (br, 2746H), 3.41 (br, 92H), 3.36 - 2.90 (m, 121H), 2.20 - 0.86 (m, 645H). Synthesis of Compound 101B04

[0368]

Chemical Structure

[0369] 101B03 (8.69 g) was dissolved in 45 ml of dichloromethane, 22 ml of TFA was added, protected with nitrogen, and stirred overnight at room temperature. The solvent was removed by spin distillation to obtain 6.178 g of a transparent viscous substance 101B04.

[0370] 1 H NMR (400 MHz, Deuterium Oxide) δ 4.09 (br, 32H), 3.91 (br, 91H), 3.57 (br, 2658H), 3.25 (br, 89H), 3.20 ‐ 2.89 (m, 120H), 1.93 ‐ 1.05 (m, 353H). Synthesis of Compound 101B05

[0371]

Chemical Structure

[0372] 101B04 (6.178 g) was dissolved in 21 ml of DMF, DIPEA (3.01 ml) was added, diglycolic anhydride (1.35 g) was added, and stirred at room temperature. The reaction was carried out for 4 hours, diglycolic anhydride (338.5 mg) and DIPEA (754 μl) were further added, stirred overnight at room temperature, the methyl tert-butyl ether of the product was precipitated, filtered, and the cake was dried in vacuo to obtain the product 101B05.

[0373] Synthesis of Compound 101B00A

[0374]

Chemical Structure

[0375] 101B05 (0.230 g) was dissolved in 2 ml of DMF, 10109 (0.208 g, prepared in Example 11) was added, PyBop (229 mg) and DIPEA (0.115 ml) were added, and the mixture was stirred overnight at room temperature. The product was precipitated with methyl tert-butyl ether, the solid was dissolved in 70 ml of methanol, 30 ml of water was added, purified by ultrafiltration, and lyophilized to obtain 310 mg of the final product (101B00A) as a red solid, with an HPLC purity of 98.8%.

[0376] Example 16: Synthesis of Compound 101B00B Synthesis of Compound 101B06

[0377]

Chemical Structure

[0378] α-[Boc-Lys(NH3 + COO - )] 30 -ε-PolyLys 30 (101B02) (prepared in Example 46) (4.76 g) was dissolved in 20 ml of water, 1014-2K (10.00 g) was dissolved in 20 ml of acetonitrile, the two solutions were mixed, triethylamine (1.39 ml) was added, and the reaction was carried out with stirring for 10 minutes. A solution of 1014-2K (20.00 g) in acetonitrile (40 ml) was added, triethylamine (2.79 ml) was added, the reaction was carried out at 25 °C for 4 hours, water was added, ultrafiltration was carried out, and the filtrate was concentrated and lyophilized to obtain 18.4 g of α-[Boc-Lys(mPEG 2k )] 30 -ε-PolyLys 30 as a white powdery product (101B06).

[0379] 1H NMR (400 MHz, Deuterium Oxide) δ 4.25 (s, 32H), 4.09 (s, 95H), 3.93 - 3.45 (m, 5537H), 3.41 (s, 95H), 3.36 ‐ 2.90 (m, 126H), 2.20 - 0.86 (m, 645H). Synthesis of Compound 101B07

[0380]

Chem.

[0381] 101B06 (9.0 g) was dissolved in dichloromethane (90 ml), TFA (30 ml) was added, and the mixture was stirred at room temperature for 6 hours. The solvent was removed by spin distillation, and dichloromethane was dried under vacuum to obtain 8.84 g of white solid 101B07 (97.6%).

[0382] 1 H NMR (400 MHz, Deuterium Oxide) δ 4.23 (br, 35H), 4.04 (br, 94H), 3.70 (br, 5255H), 3.38 (br, 92H), 3.25 (m, 128H), 2.10 ‐ 1.11 (m, 376H). Synthesis of Compound 101B08

[0383]

Chem.

[0384] 101B07 (1.50 g) was dissolved in 6 ml of DMF, diglycolic anhydride (0.25 g) was added, DIPEA (0.66 ml) was added, and the mixture was stirred at room temperature. Diethylene glycol anhydride (189 mg) and DIPEA (528 μl) were added, and the mixture was stirred overnight at room temperature. The product was precipitated with MTBE, filtered, and the cake was dried under vacuum to obtain white solid 101B08 (DIPEA salt).

[0385] Synthesis of Compound 101B00B

[0386]

Chem.

[0387] 101B08 (0.43 g), 10109 (0.24 g) and PyBop (0.13 g) were dissolved in 4 ml of DMF, DIPEA (0.13 ml) was added, and the mixture was stirred overnight at room temperature. 30 ml of MTBE was added to the reaction solution to precipitate, centrifuged, the dichloromethane and the precipitate were dried under vacuum, dissolved in 70 ml of methanol, 30 ml of water was added, purified by ultrafiltration, and freeze-dried to obtain 480 mg of the final product (101B00B) as a red solid, HPLC purity: 98.0%.

[0388] Example 17: Measurement of DOX Coupling in Polymers 101A1601, 101A1602, 101B00A, 101B00B Papain was weighed and added to PBS to form a 6.25 mg / ml PBS solution.

[0389] Composition of polymer solution: Four types of polymers 101A1601, 101A1602, 101B00A, 101B00B were weighed into a round-bottom flask with a known mass according to the following table, 2 ml of methanol was added to dissolve the polymer, a certain amount of water was added and spin-distilled to remove methanol, the mass of the remaining liquid was weighed by the differential method, and water was added so that the concentration of the polymer was 25 mg / ml.

[0390]

Table 4

[0391] 0.20 ml of the polymer solution and 1.80 ml of the papain PBS solution were taken, the final concentration of the polymer was 2.50 mg / ml, and it was shaken in a water bath at 37 °C for 72 hours. After cooling to room temperature, 0.10 ml was pipetted and added to 0.90 ml of DMSO. The peak area of DOX after enzymatic digestion was characterized by HPLC, and the amount of DOX coupling was calculated using the standard curve of DOX-HCl (converted to DOX·HCl).

[0392] Among these, 101B00B, 101A1601, and 101A1602 could not be completely digested even after 72 hours. Therefore, using 101B00A as the standard sample, the amount of DOX coupling of these 3 polymers was calculated.

[0393]

Table 5

[0394] Example 18: Synthesis of Compound 101B00C: Synthesis of Compound 10110

[0395]

Chemical Structure

[0396] SN-38 (28.87 g) was added to a 2 L round-bottom flask, 840 mL of THF was added to obtain a yellow suspension, and after cooling in an ice bath for 10 minutes, p-nitrophenyl chloroformate (18.09 g) was added and protected with nitrogen. DIPEA (20.12 mL) was added dropwise. The ice bath was removed, and the reaction was stirred at room temperature for 2 hours. THF was removed by spin distillation, 500 mL of ethyl acetate was added, shaken for 2 hours, and filtered. 42.35 g of pale yellow powder pNP-10-O-SN-38 (10110) was obtained by vacuum drying. LCMS (m / z): [M+H] + = 557.8 Synthesis of Compound 10111

[0397]

Chemical Structure

[0398] pNP-10-O-SN-38(10110) (41.01 g) was added to 180 mL of dichloromethane to obtain a yellow paste. A dichloromethane solution of N-Boc-piperazine (27.40 g / 90 mL) was added, and the mixture was ice-bathed for 10 minutes. DMAP (8.99 g) was added in several portions, and the reaction was stirred at room temperature overnight for 15 hours. Dichloromethane was removed by spin distillation, 180 mL of methanol was added to 1100 mL of MTBE, filtered, and dried under vacuum to obtain 33.89 g of the product N-Boc-piperazine-10-O-SN-38(10111) in 87.2% yield. LCMS(m / z): [M+H] + = 604.9 Synthesis of Compound 10112

[0399]

Chemical Structure

[0400] N-Boc-piperazine-10-O-SN-38(10111) (38.79 g) was dissolved in 150 mL of dichloromethane, 130 mL of TFA was added, stirred at room temperature for 3 hours, concentrated by distillation, 1.4 L of MTBE was added, filtered, and the cake was dried under vacuum to obtain 37.99 g of the product TFA-piperazine-10-O-SN-38(10112) in 95.7% yield. LCMS(m / z): [M+H] + = 504.9; [M+2H] + / 2 = 253.0 Synthesis of Compound 10113

[0401]

Chemical Structure

[0402] TF A - piperazine - 10 - O - SN - 38 (10112) (2.71 g) and Fmoc - Val - Cit - PABC - pNP (10107) (3.36 g) were dissolved in DMF (34 mL), DIPEA (2.29 mL) was added, and the reaction was carried out at room temperature for 3 hours; 40 mL of MTBE was added, filtered, and by vacuum drying, 4.71 g of the product Fmoc - Val - Cit - PABC - piperazine - 10 - O - SN - 38 (10113) was obtained with a yield of 95.0%. LCMS (m / z): [M + H] + = 1131.8; [M + 2H] + / 2 = 566.4 Synthesis of Compound 10114

[0403]

Chemical Structure

[0404] Fmoc - Val - Cit - PABC - piperazine - 10 - O - SN - 38 (10113) (4.71 g) was dissolved in 10% piperidine / DMF (32 mL), reacted at room temperature for 10 minutes, 210 mL of MTBE was added, filtered, and a pale yellow filter cake was obtained, which was dried under vacuum. 3.58 g of the product 10114 was obtained with a yield of 94.6%. LCMS (m / z): [M + H] + = 909.9; [M + 2H] + / 2 = 455.6 Synthesis of Compound 101B00C:

[0405]

Chemical Structure

[0406] 0.423 g of 101B08 and 0.231 g of 10114 were dissolved in 4 ml of DMF, and 0.176 g of PyBop was added. 0.133 ml of DIPEA was added, and the mixture was stirred at room temperature for 6 hours. 30 ml of MTBE was added to precipitate, and centrifuged. 3 ml of methanol / 3 ml of DCM was added to redissolve the precipitate, and then 30 ml of MTBE was added again to precipitate, followed by centrifugation. The crude product of 101B00C was obtained by vacuum drying. 4 ml of DMSO was added to the crude product, dissolved with stirring, 6 ml of methanol was added, filtered through a 0.45 μm membrane, 10 ml of water was added, and ultrafiltration was performed. It was diluted with 50% ACN / H2O each time and freeze-dried to obtain 392 mg of solid with an HPLC purity of 95%.

[0407] Example 19: Synthesis of Compound 101B00D: Synthesis of Compound 10117-2k

[0408]

Chemical Structure

[0409] mPEG (50.0 g, average molecular weight about 2000) was dissolved in 250 ml of DCM, p-nitrophenyl chloroformate (pNP-Cl, 15.12 g) was added, and triethylamine (11.9 ml) was added. The mixture was stirred overnight under nitrogen protection. The reaction solution was concentrated by spin distillation, 250 mL of MTBE was added to precipitate to 300 mL, filtered, and vacuum dried to obtain the product (10117-2k), 46.89 g, off-white, with a yield of 86.6%.

[0410] 1 H NMR (400 MHz, Chloroform-d) δ 8.28 (d, J = 9.2 Hz, 2H), 7.40 (d, J = 9.2 Hz, 2H), 4.43 (s, 2H), 3.64 (s, 172H), 3.38 (s, 3H). Synthesis of Compound 101B09

[0411] [Chemical]

[0412] 101B02 (0.32 g) was dissolved in 1.44 ml of water, 10117-2k (2.88 g) was dissolved in 4.15 ml of acetonitrile, the two solutions were mixed, 0.50 ml of DIPEA was added, and the mixture was stirred overnight at room temperature. The reaction solution was added to water, extracted with DCM, dried over anhydrous magnesium sulfate, filtered, the filtrate was concentrated, precipitated with methyl tert-butyl ether, and dried under vacuum to obtain 1.51 g of product 101B09, a pale yellow solid (87.5%).

[0413] Synthesis of Compound 101B10

[0414] [Chemical]

[0415] 101B09 (1.51 g) was dissolved in 20 ml of DCM, TFA (10 ml) was added, and the mixture was stirred overnight at room temperature. Most of the solvent was removed by spin distillation, the product was precipitated with methyl tert-butyl ether, and 101B10 was obtained by vacuum drying and used directly in the next step.

[0416] Synthesis of Compound 101B12

[0417] [Chemical]

[0418] 101B10 (5.17 g, 2.13 mmol) was dissolved in 20 ml of DMF. Succinic anhydride (320 mg, 3.2 mmol) and triethylamine (0.89 ml, 6.4 mmol) were added. The reaction was stirred at room temperature for 4 hours, precipitated with methyl tert-butyl ether, centrifuged, and dried under vacuum to obtain 5.01 g (94%) of 101B12 as a white powder.

[0419] Synthesis of Compound 101B00D

[0420]

Chem.

[0421] 0.709 g of 101B12 and 0.386 g of 10114 were dissolved in 6 ml of DMF, 0.220 g of PyBop was added, 0.221 ml of DIPEA was added, and the mixture was stirred overnight at room temperature. 40 ml of MTBE precipitation was added, centrifuged, a small amount of precipitated methanol / water was taken and dissolved, 3 ml of methanol / 3 ml of DCM suspension was precipitated, followed by precipitation with MTBE, centrifuged, a small amount of precipitated methanol was taken and dissolved, and vacuum dried to precipitate, obtaining a crude product.

[0422] The crude product was dissolved in an acetonitrile / water (1:1) mixed solvent, filtered through a 0.45 μm filter membrane, and ultrafiltered at 30 kDa. As a result, the purity of 101B00D increased from 72.4% to 98.3% (at 210 nm).

[0423] Example 20: Synthesis of Compound 101B00E Synthesis of Compound 101B13

[0424]

Chem.

[0425] 101B07 (27.37 g) was dissolved in DMF (104 ml), succinic anhydride (2.373 g) was added, DIPEA (8.174 ml) was added, and the mixture was stirred at room temperature for 4 hours. A small amount of methyl tert-butyl ether precipitate was taken, reacted with ninhydrin at 100 °C for 3 minutes, and became colorless. 300 ml of MTBE precipitate was added to obtain a mucus, which was centrifuged, the supernatant was diluted, and GPC characterization without product was performed. The viscous substance was dissolved in 100 ml of ACN, 350 ml of MTBE was slowly added to precipitate the product, and the solution was removed. The gel-like precipitate was dissolved in 100 ml of ACN, 400 ml of MTBE was slowly added to precipitate the product, which was filtered, and the product was vacuum dried to obtain 21.840 g (8.7 mmol, 76%). The measured succinic acid content was 218 ppm.

[0426] 1 H NMR (400 MHz, Deuterium Oxide) δ 4.16 (br, 61H), 4.00 (br, 63H), 3.64 (br, 5113H), 3.32 (br, 91H), 3.27 ‐ 2.84 (m, 124H), 2.80 ‐ 2.30 (br, 127H), 2.14 ‐ 0.71 (br, 366H). Synthesis of Compound 101B00E

[0427]

Chemical Structure

[0428] 101B13 (4.873 g, 1.94 mmol) was dissolved in 40 ml of DMF, 10114 (2.648 g, 2.91 mmol) and PyBop (2.027 g, 3.88 mmol) were added, DIPEA (1.52 ml, 8.74 mmol) was added, and the mixture was stirred at room temperature for 4.5 hours. The product was precipitated with methyl tert-butyl ether and purified by ultrafiltration with MeOH / water (50 / 50, v / v), and the concentrate was lyophilized to obtain 5.51 g of the final product 101B00E, a pale yellow cotton-like solid.

[0429] HPLC analysis method D: 101B00E: RT = 12.602 minutes Measurement of SN-38 coupling of compound 101B00E Papain was weighed and added to PBS to form a 10.0 mg / ml PBS solution.

[0430] Lyophilized 101B00E (ε-PolyLys 30 -[Lys(α-SA-VC-PABC-piperazine-10-O-SN-38, ε-mPEG 2k )] 30 ) 38.39 mg was weighed, and 3.839 ml of deionized water was added to obtain a 10.0 mg / ml polymer solution.

[0431] 0.500 ml of the polymer solution and 0.500 ml of the papain PBS solution were taken, the final concentration of the polymer was 5.00 mg / ml, and it was shaken in a 37°C water bath for 2 hours. It was cooled to room temperature, 0.200 ml was pipetted, 1.800 ml of DMSO was added, the peak area of piperazine-10-O-SN-38 after enzymatic digestion was characterized by HPLC, and when the concentration of TFA-piperazine-10-O-SN-38 was calculated using the standard curve of TFA-piperazine-10-O-SN-38, it was 841.1 μg / ml. After conversion, the concentration of SN-38 after enzymatic hydrolysis of 5.00 mg / ml polymer was 506.4 μg / ml, and the coupling amount of SN-38 in ε-PolyLys 30 -[Lys(α-SA-VC-PABC-piperazine-10-O-SN-38, ε-mPEG 2k )] 30 was calculated to be 10.1 wt.%.

[0432] Example 21: Synthesis of compound 101B00G Synthesis of compound 10111A

[0433]

Chemical formula

[0434] Dichloromethane (5 mL) was added to pNP-10-O-SN-38 (1.30 g) to obtain a yellow paste. Boc-DMEDA(101M15) (0.87 g) and DIPEA (406 μL) were added, and the solution was gradually dissolved, clarified, and further turned light brown. After 3 hours of reaction, the solvent was removed by spin drying. After column purification, 1.0749 g (76.0%) of the product Boc-DMEDA-10-O-SN-38(10111A) was obtained. LCMS (m / z0: [M+H] + = 607.2 1 HNMR (400 MHz, CDCl3): δ=8.22 (d, 1H, J = 8.8Hz), 7.90 - 7.84 (m, 1H), 7.64 (s, 1H), 7.61 - 7.59 (m, 1H), 5.75 (d, 1H, J = 16.4Hz), 5.33 - 5.26 (m, 4H), 3.75 (s, 1H), 3.65 - 3.54 (m, 3H), 3.26 - 2.78 (m, 9H), 1.94 - 1.85 (m, 2H), 1.48 - 1.40 (m, 10H), 1.28 - 1.25 (m, 3H), 1.06 - 1.02 (m, 3H). Synthesis of Compound 10112A

[0435]

Chemical Structure

[0436] Boc-DMEDA-10-O-SN-38 (0.7429 g) was dissolved in 5 mL of dichloromethane, becoming turbid. 2 mL of TFA was added, and the reaction mixture was stirred at room temperature for 1 hour, concentrated, and the crude product TFA-DMEDA-10-O-SN-38(10112A) was used directly in the next reaction. LCMS (m / z): [M+H] + = 507.1 Synthesis of Compound 10113A

[0437]

Chemical Structure

[0438] TFA-DMEDA-10-O-SN-38 (10112A) (732.5 mg) and Fmoc-Val-Cit-PABC-pNP (10107) (905.1 mg) were dissolved in DMF (9 mL), DIPEA (620 μl) was added, and the mixture was reacted at room temperature for 3 hours. 90 mL of MTBE was added, centrifuged, and by vacuum drying, 1.1213 g (83.8%) of Fmoc-Val-Cit-PABC-DMEDA-10-O-SN-38 (10113A) was obtained. LCMS (m / z): [M+H] + / 2 = 567.75 Synthesis of Compound 10114A

[0439]

Chemical Structure

[0440] Fmoc-Val-Cit-PABC-DMEDA-10-O-SN-38 (10113A) (1.1213 g) was added to a 10 mL round-bottom flask, 5 mL of DMF was added to dissolve it, 0.56 mL of DEA was added, and the mixture was stirred for 20 minutes. 40 mL of MTBE was added and centrifuged. The precipitate was dissolved in 4 mL of MeOH, precipitated with 40 mL of MTBE, and centrifuged to obtain 0.7128 g of NH2-Val-Cit-PABC-DMEDA-10-O-SN-38 (10114A) as a pale yellow solid with a yield of 79.1%. LCMS (m / z): [M+H] + = 912; [M+2H] + / 2 = 456.7 Synthesis of Compound 101B00G

[0441]

Chemical Structure

[0442] 101B13 (0.705 g), 10114A (0.384 g), and PyBop (0.293 g) were dissolved in 6 ml of DMF, DIPEA was added, and the mixture was stirred at room temperature for 4 hours. MTBE was added to precipitate, and the precipitate was added to 60 ml of methanol and 70 ml of water, purified by ultrafiltration, and freeze-dried to obtain 0.832 g of the final product 101B00G, a pale yellow cotton-like product.

[0443] HPLC analysis method D, RT = 13.824 minutes.

[0444] 1 H NMR (400 MHz, DMSO-d6) δ 8.12 (br, 62H), 7.89 (br, 66H), 7.61 (br, 132H), 7.28 (br, 98H), 5.35 (br, 112H), 4.98 (br, 70H), 4.26 (br, 150H), 3.84 (br, 77H), 3.50 (br, 6237H), 3.23 (br, 90H), 3.18 ‐ 2.74 (m, 374H), 1.34 (m, 899H). Measurement of the amount of SN-38 coupling in 101B00G Papain was weighed and added to PBS to form a 10.0 mg / ml PBS solution.

[0445] Freeze-dried ε-PolyLys 30 -[Lys(α-SA-VC-PABC-DMEDA-10-O-SN-38, ε-mPEG 2k )] 30 (101B00G) was weighed at 35.12 mg, 3.510 ml of deionized water was added to obtain a 10.0 mg / ml polymer solution.

[0446] 0.50 ml of the polymer solution and 0.50 ml of the papain PBS solution were taken. The final concentration of the polymer was 5.00 mg / ml, and it was shaken in a water bath at 37 °C for 2 hours. After cooling to room temperature, 0.20 ml was pipetted out, 1.800 ml of DMSO was added, and it was characterized by HPLC. By calculating the SN-38 concentration after polymer enzymatic hydrolysis according to the SN-38 standard curve, the SN-38 concentration after polymer enzymatic hydrolysis at 5.00 mg / ml was 472.8 μg / ml, and the SN-38 coupling amount in 101B00G was calculated to be 9.5 wt.%.

[0447] Example 22: Synthesis of Compound 101B00H Synthesis of Compound 10111B

[0448]

Chemical formula

[0449] Dichloromethane (5 mL) was added to pNP-10-O-SN-38 (10110) (1.30 g), 1-BOC-3-methylaminoazetidine (101M16) (0.87 g) and DIPEA (406 μL) were added, and the reaction was carried out at room temperature for 3.5 hours. It was concentrated by spin distillation, 5 mL of MeOH was added, 80 mL of MTBE was added, centrifuged and precipitated, and dried in vacuo to obtain 1.24 g of the product 1-Boc-Az-10-O-SN-38 (10111B) and the yield. LCMS (m / z): [M+H] + = 605 Synthesis of Compound 10112B

[0450]

Chemical formula

[0451] 1-Boc-Az-10-O-SN-38 (10111B) (1.00 g) was dissolved in 10 mL of dichloromethane, 3 mL of TFA was added, and the mixture was stirred at room temperature for 1 hour. It was concentrated by spin distillation, 80 mL of MTBE was added, centrifuged, and dried under vacuum. The product Az-10-O-SN-38 (10112B) 0.97 g, 94.4% was obtained. LCMS (m / z): [M+H] + =505.1 Synthesis of Compound 10113B

[0452]

Chemical Structure

[0453] Az-10-O-SN-38 (10112B) (966.6 mg) and Fmoc-Val-Cit-PABC-pNP (10107) (1.1743) were dissolved in 12 mL of DMF, DIPEA (816 μL) was added, and the mixture was reacted at room temperature for 3 hours. 80 mL of MTBE was added, centrifuged, and dried under vacuum. The product Fmoc-Val-Cit-PABC-Az-10-O-SN-38 (10113B) 1.60 g, 90.7% was obtained. LCMS (m / z): [M+2H] + / 2 =566.4 Synthesis of Compound 10114B

[0454]

Chemical Structure

[0455] Fmoc-Val-Cit-PABC-Az-10-O-SN-38 (10113B) (1.6040 g) was dissolved in 10 mL of DMF, 1.2 mL of DEA was added to obtain a light yellow solution. After stirring for 15 minutes, 80 mL of MTBE was added, centrifuged, and the precipitate was dried under vacuum. The product NH2-Val-Cit-PABC-Az-10-O-SN-38 (10114B) 1.2630 g, 97.9% was obtained. LCMS (m / z): [M+H] + =910.2; [M+2H] + / 2 =455.7 Measurement of Compound 101B00H

[0456] [Chemistry]

[0457] 101B13 (0.70 g), 10114B (0.38 g), and PyBop (0.29 g) were dissolved in DMF (6 ml), DIPEA (0.22 ml) was added, and the mixture was stirred at room temperature for 3 hours. The complete reaction of the components was characterized by HPLC. MTBE was added to precipitate, and the precipitate was added to 5 ml of methanol and 5 ml of water, purified by ultrafiltration, and freeze-dried to obtain 881 mg of the final product 101B00H, a pale yellow cottony product. HPLC analysis method D, RT = 13.716 minutes.

[0458] Measurement of the Amount of SN-38 Coupling in 101B00H Papain was weighed and added to PBS to form a 10.0 mg / ml PBS solution.

[0459] Freeze-dried ε-PolyLys 30 -[Lys(α-SA-VC-PABC-Az-10-O-SN-38, ε-mPEG 2k )] 30 (101B00H) 29.77 mg was weighed, 0.149 ml of DMSO was added and heated to dissolve, and 2.83 ml of deionized water was added to obtain a 10.0 mg / ml polymer solution.

[0460] 0.50 ml of the polymer solution and 0.50 ml of the papain PBS solution were taken, the final concentration of the polymer was 5.00 mg / ml, and the mixture was shaken in a 37°C water bath for 2 hours. It was cooled to room temperature, 0.20 ml was pipetted and added to 1.80 ml of DMSO, characterized by HPLC, and when the concentration of 10112B (Az-10-O-SN-38 trifluoroacetate) after polymer digestion was calculated to be 855.2 μg / ml using the 10112B (Az-10-O-SN-38 trifluoroacetate) standard quartile diagram, and the concentration of SN-38 was converted to 514.8 μg / ml, ε-PolyLys 30-[Lys(α-SA-VC-PABC-Az-10-O-SN-38, ε-mPEG 2k )] 30 (101B00H), the SN-38 coupling amount was calculated to be 10.3 wt.%.

[0461] Example 23: Synthesis of Compound 101B00I Synthesis of Compound 10121

[0462]

Chemical Structure

[0463] 101M17 (20.0 g) was dissolved in 200 ml of MeOH, concentrated sulfuric acid (1.17 g) was added, and the mixture was refluxed at 75 °C for 2 hours under nitrogen protection. The reaction solution was cooled to room temperature, spin-dried, EA and saturated NaHCO3 were added, and the organic phase was dried over anhydrous sodium sulfate. After filtration and spin-drying, 18.48 g of the product 10121, an orange-red viscous oil, was obtained with a yield of 85.3%.

[0464] Synthesis of Compound 10122

[0465]

Chemical Structure

[0466] 10121 was dissolved in 50 mL of chloroform, acetone dimethanol (22.04 mL) and p-TsOH-H2O (0.84 g) were added, and the reaction solution was refluxed at 70 °C overnight. Chloroform was removed by spin-drying, 50 mL of DCM was added, the reaction solution was neutralized by adding 100 mL of NaHCO3, partitioned into the liquid phase, the aqueous phase was extracted with DCM, dried over anhydrous sodium sulfate, and dried by filtration and spin-drying. Purification by column chromatography gave 2.21 g of the product 10122 with a yield of 36.2%.

[0467] Synthesis of Compound 10123

[0468]

Chem.

[0469] 10122 (2.21 g) was dissolved in MeOH, an aqueous LiOH solution was added, and the mixture was stirred overnight at room temperature. MeOH was removed by spin distillation, the pH was adjusted to 5 - 6 with 1M hydrochloric acid, extracted with DCM, dried over anhydrous sodium sulfate, spin-dried, and dried under vacuum to obtain 1.76 g of product 10123, in a yield of 84.9%, as pale yellow crystals.

[0470] 1 H NMR (400 MHz, Chloroform-d) δ 6.70 (d, J = 5.7 Hz, 3H), 3.56 (s, 2H), 1.69 (s, 6H). Synthesis of Compound 10124

[0471]

Chem.

[0472] 5 ml of DCM was added to 10123 (0.500 g) and NHS (0.415 g), and EDCl (0.691 g) was added. The mixture was stirred and reacted at room temperature for 2 hours. A small amount was taken, washed with water, diluted with acetonitrile, ESI-MS: [M+NH4] + = 323. The organic phase was washed twice with water, dried over anhydrous sodium sulfate, spin-dried to obtain 834 mg of 10124, which was directly used in the next step.

[0473] Synthesis of Compound 101B14

[0474]

Chem.

[0475] 101B10 (3.70 g) was dissolved in 8 ml of DMF, DIPEA (0.83 ml) was added, 10124 (0.72 g) was dissolved in 5 ml of DMF, added to the above, and stirred overnight at room temperature. MTBE was added to precipitate the reaction solution, and vacuum dried to obtain 101B14, 3.17 g of a pale yellow solid (83.3%).

[0476] 1 H NMR (400 MHz, DMSO-d6) δ 6.69 (57H), 6.50 (9H), 4.03 (s, 126H), 3.51 (5240H), 3.24 (90H), 2.90 (160H), 1.59 (351H). Synthesis of Compound 101B15

[0477]

Chemical Structure

[0478] 101B14 (1.00 g) was dissolved in 5 ml of chloroform, 1.5 ml of TFA was added, 147 μl of water was added, and heated to 50 °C under nitrogen protection with stirring overnight. The reaction solution was precipitated with MTBE and vacuum dried to obtain 857 mg (87.0%) of the product 101B105.

[0479] Synthesis of Compound 101B00I

[0480]

Chemical Structure

[0481] Bortezomib (13.54 mg) was weighed and dissolved in 271 μl of DMSO. 101B15 (168.7 mg) was dissolved in 5 ml of phosphate buffer (0.01 M, pH = 8.1), the DMSO solution of bortezomib was added, purified by ultrafiltration, and lyophilized to obtain the final product, the final product of 7.2 wt.% bortezomib.

[0482] Example 24: Synthesis of Compound 101B00J-S Synthesis of Compound 101B00J-S

[0483]

Chemical Structure

[0484] 101B07 (0.850 g) was dissolved in 5 ml of anhydrous DMF. 10303 (0.570 g) was dissolved in 5 ml of anhydrous DMF, the two solutions were mixed, and DIPEA was added. The reaction solution was stirred at room temperature overnight, MTBE was added to precipitate, the precipitate was dissolved in 100 ml of methanol / water and purified by ultrafiltration, and freeze-dried to obtain 681 mg of the final product 101B00J-S, a white solid.

[0485] HPLC Analysis Method A, RT = 8.870 minutes.

[0486] Measurement of the Amount of PTX Coupling in 100B00J-S The amount of PTX coupling was quantified using 10302 as a standard.

[0487] 101B00J-S: 0.807 mg / ml, Area = 4009.3, corresponding to 0.195 mg / ml of 10302, 10302 coupling amount: 24.2%.

[0488] 10302 molecular weight: 986.05, PTX molecular weight 853.92, PTX coupling amount: 20.9%.

[0489] The concentration of 101B00J-S was 39.98 mg / ml, and the PTX concentration was 8.36 mg / ml.

[0490] 516.2 mg of 101B00J-S was dissolved in 10.20 g of water to make the polymer concentration 50.57 mg / ml. 100 μl was taken, 7.90 ml of water was added to make the polymer concentration 0.632 mg / ml, and this was filtered and characterized by HPLC: Area = 3399.2, corresponding 10302 was 0.166 mg / ml 10302 Standard curve: y = 20421x + 10.234 The stock solution corresponds to 11.5 mg / ml PTX Example 25: Synthesis of Compound 101B00J-V Synthesis of Compound 10401

[0491]

Chem.

[0492] Vinblastine (1.00 g) and 1,4-dioxane-2,6-dione (0.28 g) were added to a flask. After purging with nitrogen and evacuating to vacuum, a mixture of DIPEA (1.00 ml) and 50 ml of anhydrous dichloromethane was added. The reaction was carried out at room temperature for 5 hours. It was confirmed by TLC that the raw materials had basically reacted completely. The mixture was washed with 0.5 M hydrochloric acid, washed with water, dried over anhydrous sodium sulfate, spin-dried, and purified by column chromatography to obtain 0.82 g of solid product 10401.

[0493] Synthesis of Compound 10402

[0494]

Chem.

[0495] 10401 (0.50 g), NHS (0.095 g), and EDCl (0.15 g) were added to a flask, 10 ml of dichloromethane was added, and the mixture was stirred overnight at room temperature. The organic phase was washed twice with water, dried over anhydrous sodium sulfate, spin-dried, and dried under vacuum to obtain 0.55 g of solid product 10402, which was directly used in the next reaction.

[0496] Synthesis of Compound 101B00J-V

[0497]

Chem.

[0498] 101B07 (0.85 g) was dissolved in 5 ml of anhydrous DMF. 10402 (0.54 g) was dissolved in 5 ml of anhydrous DMF, the two solutions were mixed, and DIPEA was added. The reaction solution was stirred overnight at room temperature, MTBE was added to precipitate, the precipitate was dissolved in 100 ml of methanol / water and purified by ultrafiltration, and freeze-dried to obtain 721 mg of the final product 101B00J-V, a white solid.

[0499] Example 26: Synthesis of Compound 101B00J-Pt

[0500]

Chemical Structure

[0501] 101B07 (0.5 g) was dissolved in 5 ml of anhydrous DMF. 10502 (0.44 g, synthesized according to the same method in the literature, Can. J. Chem. 1993, 71(6)896) was dissolved in 5 ml of anhydrous DMF, the two solutions were mixed, and DIPEA was added. The reaction solution was stirred overnight at room temperature, MTBE was added to precipitate, the precipitate was dissolved in 100 ml of methanol / water and purified by ultrafiltration, and freeze-dried to obtain 389 mg of the final product 101B00J-Pt.

[0502] Example 27: Synthesis of Compound 101B00F Synthesis of Compound 10118

[0503]

Chemical Structure

[0504] Melphalan (101M14) (500.0 mg) was dispersed in 16 ml of methanol, cooled in an ice bath, 0.48 ml of SOCl2 was added dropwise, the solvent of the mixture was heated to 65 °C, and the reaction was refluxed for 18 hours. The solvent was removed by spin distillation and dried under vacuum. 523 mg of the crude product Melphalan-OMe (10118) was used directly in the next step. LCMS (m / z): [M+H] + = 319.0 Synthesis of Compound 10119

[0505]

Chem.

[0506] Melphalan-OMe·HCl (10118) (381.3 mg) and Fmoc-Val-Cit-PABC-pNP (10107) (805.6 mg) were dissolved in 5 mL of DMF, DIPEA (560 μL) was added, and the reaction was carried out at room temperature for 4 hours. 80 mL of MTBE was added, centrifuged, and the precipitate was dried under vacuum. The product Fmoc-Val-Cit-PABC-Melphalan-OMe (10119) 0.95 g, 95.9% was obtained. LCMS (m / z): [M+H] + = 946.2 Synthesis of Compound 10120

[0507]

Chem.

[0508] Fmoc-Val-Cit-PABC-Melphalan-OMe (10119) (1.39 g) was dissolved in 5 mL of DMF, 0.7 mL of DEA was added, and the reaction was carried out at room temperature for 15 minutes. 100 mL of MTBE was added, centrifuged, and dried under vacuum to obtain the product NH2-Val-Cit-PABC-Melphalan-OMe (10120) 1.05 g, yield 98.8%. LCMS (m / z): [M+H] + =724; [M+2H] + / 2 =362.6 Synthesis of Compound 101B00F

[0509]

Chem.

[0510] 101B13 (0.840 g), 10120 (0.364 g), and PyBop (0.348 g) were dissolved in 9 ml of DMF and stirred at room temperature for 3 hours. MTBE was precipitated, and the crude product of the precipitate was dissolved in 50% MeOH / H2O ultrafiltration purification and freeze-dried to obtain the final product (101B00F), 955.5 mg, a brown solid.

[0511] HPLC analysis method C, RT = 9.093 minutes.

[0512] Example 28: Synthesis of Compound 101B00K Compound 101B00K was synthesized according to the same method as in Examples 15 - 17 and Example 70:

[0513]

Chemical Structure

[0514] Example 29: Synthesis of Compound 101B00L Compound 101B00L was synthesized according to the same method as in Examples 15 - 17 and Example 70:

[0515]

Chemical Structure

[0516] Example 30: Synthesis of Compound 101B00M Compound 101B00M was synthesized according to the same method as in Examples 15 - 17 and Example 70:

[0517]

Chemical Structure

[0518] Example 30: Synthesis of Compound 101B00N Compound 101B00N was synthesized according to the same method as in Examples 15 - 17 and Example 70:

[0519]

Chemical Structure

[0520] Example 31: Synthesis of Compound 101B00O 101B00O was synthesized according to the same method as in Examples 15 to 17 and Example 70:

[0521]

Chemical formula

[0522] Example 32: Synthesis of Compound 101B00P The synthesis of 21209 refers to the synthesis process of Compound 3c mentioned in the literature European Journal of Medicinal Chemistry, 2021, 216, 113355 Synthesis of Compound 101B00P

[0523]

Chemical formula

[0524] 101B08 (1.500 g) and 21209 (0.753 g) were dissolved in 7.5 ml of DMF, 0.625 g of PyBop was added, 0.471 ml of DIPEA was added, and the mixture was stirred at room temperature for 24 hours.

[0525] 30 ml of MTBE was added to cause precipitation, and the mixture was centrifuged. 3 ml of methanol / 3 ml of DCM was added to dissolve it, and then 30 ml of MTBE was continuously added to cause precipitation, and the mixture was centrifuged. A small amount of precipitated methanol was taken and dissolved. The precipitate was dried under vacuum to obtain a crude product.

[0526] The crude product was dissolved in 20 ml of methanol, added to 150 ml of 50% MeOH, purified by ultrafiltration, the purified solution was concentrated to ~26 ml by spin distillation, partitioned, and freeze-dried to obtain 1.707 g of a white solid (structural unit molecular weight 3242, 0.533 mmol, 88.8%, HPLC 99.6%).

[0527] 1H NMR (400 MHz, DMSO-d6) δ 7.73 ‐ 7.41 (m, 61H), 7.25 (s, 65H), 7.11 (d, J = 8.5 Hz, 33H), 6.96 (d, J = 12.2 Hz, 67H), 6.64 ‐ 6.35 (m, 128H), 4.97 (s, 65H), 4.27 (d, J = 92.8 Hz, 285H), 3.97 (s, 63H), 3.71 (s, 5400H), 3.35 (s, 84H), 3.27 ‐ 2.75 (m, 391H), 2.28 ‐ 1.14 (m, 549H), 0.95 (t, J = 8.6 Hz, 203H). Measurement of CA4 coupling of 101B00P 15 - 20 mg of the lyophilized product of 101B00P (recorded as mass m A is placed in a well-sealed glass sample (plastic centrifuge tubes are prohibited to avoid concentration bias due to liquid leakage), water is added and sealed, and the amount of water added is 1.5 - 2.0 ml so that the final concentration of 101B00P is 10 mg / ml, and the mass m AW is recorded, and it is shaken at 37 °C for 20 minutes to dissolve and becomes a colorless and transparent liquid.

[0528] Composition of the buffer solution Sodium dihydrogen phosphate: 352 mM Disodium hydrogen phosphate: 48 mM EDTA·2Na: 4.0 mM L-Cysteine hydrochloride: 8.0 mM Brij 35: 0.1% (v / v) Adjust the pH to 6.0 with 1 M HCl or 1 M NaOH Composition of the buffer solution The buffer solution with a papain B concentration of 10 mg / ml was prepared as described above.

[0529]

Table 6

[0530] Tear a 1.5 ml vial with a 10000 mm balance, and pipette 200 μl of 101B00P aqueous solution (mass m A1 ) into a 1.5 ml vial with a 200 μl pipette gun, add 200 μl of papain B solution, cap and seal it, and vortex at 1000 rpm for 30 seconds.

[0531] Decompose this solution with a shaker at 37 °C for 20 hours or more.

[0532] Take it out from the shaker, cool it to room temperature, then transfer the whole solution to a 10 ml volumetric flask with a 200 μl pipette gun, wash the tip of the gun and the injection bottle 3 times with 400 μl of 1% DIPEA / methanol solution, then transfer it to the volumetric flask and make up the solution to the mark (the pipette gun made up the solution to near the mark line with 200 μl) (Note: The pH of the solution after volume adjustment is 8 or higher).

[0533] The CA4 concentration was measured by HPLC after filtration [CA4] A (295 nm), and the concentration of the CA4 standard solution was 80 - 100 μg / ml (DMSO).

[0534] Calculation of the amount of CA4 coupling

[0535]

Number

[0536] The amount of CA4 coupling in the batch measured by HPLC was 9.73%, which was consistent with the theoretical coupling amount.

[0537] Example 33: Synthesis of Compound 101B00Q According to the same method as in Examples 15 - 17 and Example 70, 101B00Q was synthesized:

[0538]

Chemical formula

[0539] Example 34: Synthesis of Compound 101C00A Synthesis of Compound 101C04

[0540]

Chem.

[0541] N-Benzyloxycarbonyl-L-glutamic acid-1-tert-butyl ester was dissolved in 20 ml of DCM, 1.02 g of NHS and 1.70 g of EDCl were added, and the mixture was stirred at room temperature for 4 hours. It was washed three times with water, dried over anhydrous sodium sulfate, and spin-dried to obtain 2.849 g of a transparent foamy product (Mw = 434.45, 6.56 mmol).

[0542] Synthesis of Compound 101C05

[0543]

Chem.

[0544] ε-Polylysine hydrochloride (0.650 g) was dissolved in 0.878 ml of water and 3 ml of DMSO, 101C04 (2.573 g) was dissolved in 5.85 ml of DMSO, the two solutions were mixed, DIPEA (2.06 ml) was added, the reaction mixture was stirred at room temperature for 6 hours, 1 M NaOH and water were added, filtered, and dried in vacuo to obtain 1.709 g (96.7%) of product 101C05 as a white powder.

[0545] 1 H NMR (400 MHz, DMSO-d6) δ 8.46 ‐ 7.51 (m, 85H), 7.34 (s, 153H), 5.02 (m, 60H), 4.14 (m, 30H), 3.87 (m, 33H), 2.97 (s, 66H), 2.21 (s, 69H), 1.37 (m, 528H). Synthesis of Compound 101C06

[0546]

Chem.

[0547] 17 ml of acetic acid was placed in a 50 ml flask, 101C05 (1.68 g) was added, and the mixture was heated with stirring at 40 °C until dissolved. 840 mg of 10% palladium on carbon was added, and the mixture was stirred overnight at 30 °C under a hydrogen atmosphere. The diatomaceous earth was filtered off, the filtrate was spin distilled to remove the solvent, and the residue was dried in vacuo to obtain 1.64 g of 101C06 as a white solid.

[0548] 1 H NMR (400 MHz, Deuterium Oxide) 4.22 ‐ 3.99 (m, 30H), 3.92 (m, 29H), 3.10 (m, 104H), 2.43 (m, 60H), 2.09 (m, 55H), 1.26 (m, 566H). Synthesis of Compound 101C08

[0549]

Chem.

[0550] 101C06 (0.500 g) was dissolved in 2.68 ml of water, 10104-2k (3.724 g) was dissolved in 7 ml of acetonitrile, the two solutions were mixed, DIPEA was added, and the mixture was stirred overnight at room temperature. The acetonitrile was removed by spin distillation, water was added, the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, spin dried, and dried in vacuo to obtain 2.892 g of 101C07 as a white solid. 101C07 (2.00 g) was dissolved in 10 ml of dichloromethane, 5 ml of TFA was added, and the mixture was stirred overnight at room temperature. The solvent was removed by spin distillation, MTBE was precipitated, and 1.80 g of 101C08 was obtained as a white solid by drying in vacuo.

[0551] Synthesis of Compound 101C00A

[0552]

Chem.

[0553] 101C08 (0.66 g), 10114 (0.39 g), and PyBop (0.443 g) were dissolved in 6 ml of DMF, DIPEA (0.22 ml) was added, and the reaction mixture was stirred overnight at room temperature. The product was precipitated with 50 ml of methyl tert-butyl ether. The crude product was added to 4 ml of DMSO, dissolved with stirring, 10 ml of methanol was added, and it was purified by ultrafiltration and lyophilized to obtain 422 mg of 101C00A.

[0554] HPLC analysis method A, RT = 8.121 minutes.

[0555] Measurement of the SN-38 coupling of compound 101C00A Papain was weighed and added to PBS to form a 10.0 mg / ml PBS solution.

[0556] Lyophilized ε-PolyLys 30 -[γ-Glu-(α-CO-VC-PABC-piperazine-10-O-SN-38, α-NH-CO-mPEG 2k )] 30 30.12 mg of it was weighed, and 3.012 ml of deionized water was added to obtain a 10.0 mg / ml polymer solution.

[0557] 0.500 ml of the polymer solution and 0.500 ml of the papain PBS solution were taken, the final concentration of the polymer was 5.00 mg / ml, and it was shaken in a 37 °C water bath for 2 hours. It was cooled to room temperature, 0.200 ml was pipetted, 1.800 ml of DMSO was added, and the peak area of TFA-piperazine-10-O-SN-38 after enzymatic digestion was characterized by HPLC. Using the standard curve of TFA-piperazine-10-O-SN-38 to calculate the concentration of TFA-piperazine-10-O-SN-38, it was 876.31 μg / ml. After conversion, the concentration of SN-38 after enzymatic hydrolysis of 5.00 mg / ml polymer was 527.6 μg / ml, and ε-PolyLys 30-[γ-Glu-(α-CO-VC-PABC-piperazine-10-O-SN-38, α-NH-CO-mPEG 2k )] 30 The coupling amount of SN-38 in it was calculated to be 10.6 wt.%.

[0558] Example 35: Synthesis of Compound 101D00A Synthesis of Compound 101D07:

[0559]

Chem.

[0560] 4.944 g of N-benzyloxycarbonyl-L-glutamic acid-5-tert-butyl ester was dissolved in 50 ml of DCM, 2.53 g of NHS was added, the mixture was placed in an ice bath, 4.214 g of EDCl was added, the mixture was taken out of the ice bath and stirred overnight at room temperature. It was washed three times with water, dried over anhydrous sodium sulfate, spin-dried to obtain the product, and vacuum-dried to obtain 6.77 g of the product (15.58 mmol, 106%).

[0561] Synthesis of Compound 101D08

[0562]

Chem.

[0563] ε-Polylysine hydrochloride (1.70 g) was dissolved in 2.29 ml of water. 101D07 (6.73 g) was dissolved in 30 ml of DMSO, the two solutions were mixed, DIPEA was added, and the mixture was stirred at room temperature for 6 hours. 1M NaOH was added and the mixture was filtered, and the filter cake was vacuum-dried to obtain 4.12 g of the product 101D08, a white powder (89.2%).

[0564] 11H NMR (400 MHz, DMSO-d6) δ 8.14 ‐ 7.57 (m, 64H), 7.33 (m, 174H), 5.02 (m, 58H), 4.17 (m, 30H), 4.02 (m, 33H), 2.97 (s, 70H), 2.22 (m, 66H), 1.36 (s, 522H), 1.00 (d, J = 6.6 Hz, 38H). Synthesis of Compound 101D09

[0565]

Chem.

[0566] 39 ml of acetic acid was placed in a 50 ml flask, 101D09 (3.92 g) was added, and the mixture was stirred until dissolved. 10% palladium on carbon (0.58 g) was added, the air was evacuated, and the mixture was stirred overnight at room temperature under a hydrogen atmosphere. Celite was added, and the mixture was filtered. The filtrate was spin distilled to remove most of the solvent, precipitated with methyl tert-butyl ether, filtered, and the filter cake was dried in vacuo to obtain 2.578 g of the product 101D09.

[0567] Synthesis of Compound 101D10

[0568]

Chem.

[0569] 10 ml of DMSO was added to 101D09 (1.00 g), and the mixture was stirred until dissolved. 10104-2k (8.53 g) was added, 5 ml of DMSO and 10 ml of DMF were added, DIPEA (1.40 ml) was added, and the mixture was stirred overnight at room temperature. Precipitation with methyl tert-butyl ether, dissolution in acetonitrile, precipitation with MTBE, filtration, and drying in vacuo gave 8.15 g of the product 101D10 as a white powder. The integration from 2.165 - 2.388 ppm was fixed at 62, and the integration from 0.865 - 2.130 ppm was 514.

[0570] Synthesis of Compound 101D11

[0571]

Chem.

[0572] 101D10 (4.00 g) was dissolved in 24 ml of dichloromethane, 12 ml of TFA was added, and the mixture was stirred overnight at room temperature. The solvent was removed by spin distillation, MTBE was precipitated, ACN was dissolved, MTBE was precipitated again, and the product 101D11, 1.80 g, a white solid (60.6 %), was obtained by vacuum drying. The integral values were 62 at 2.165 - 2.388 ppm and 197 at 0.865 - 2.130 ppm, indicating that the tert-butyl ester was completely removed.

[0573] Synthesis of Compound 101D00A

[0574]

Chem.

[0575] 101D11 (0.53 g), 10114 (0.32 g) and PyBop (0.36 g) were dissolved in 5 ml of DMF. DIPEA (0.18 ml) was added and the mixture was stirred overnight at room temperature. The product was precipitated with methyl tert-butyl ether and the crude product was purified by ultrafiltration with 4 ml of DMSO and 50 ml of 50% MeOH / H2O added. The final product 101D00A, 450 mg, a pale yellow solid, was obtained by lyophilization.

[0576] HPLC Analysis Method B, RT = 7.442 min.

[0577] Measurement of SN-38 Coupling of Compound 101D00A Papain was weighed and added to PBS to form a 10.0 mg / ml PBS solution.

[0578] Lyophilized ε-PolyLys 30-[α-Glu-(γ-CO-VC-PABC-piperazine-10-O-SN-38, α-NH-CO-mPEG 2k )] 30 Weighed 28.35 mg, added 0.142 ml of DMSO, heated to dissolve the polymer, added 2.690 ml of deionized water under ultrasonic treatment conditions, and obtained a 10.0 mg / ml polymer solution.

[0579] Took 0.500 ml of the polymer solution and 0.500 ml of the papain PBS solution, with the final concentration of the polymer being 5.00 mg / ml, and oscillated in a 37 °C water bath for 2 hours. Cooled to room temperature, pipetted 0.200 ml, added 1.800 ml of DMSO, characterized the peak area of TFA-piperazine-10-O-SN-38 after enzymatic digestion by HPLC, and calculated the concentration of TFA-piperazine-10-O-SN-38 using the standard curve of TFA-piperazine-10-O-SN-38, which was 871.7 μg / ml. After conversion, the concentration of SN-38 after enzymatic hydrolysis of the 5.00 mg / ml polymer was 524.8 μg / ml, and the coupling amount of SN-38 in 30 -[α-Glu-(γ-CO-VC-PABC-piperazine-10-O-SN-38, NH-CO-mPEG 2k )] 30 ε-PolyLys was calculated to be 10.5 wt.%.

[0580] Example 36: Synthesis of Compound 101E00A Synthesis of Compound 10108-E

[0581]

Chemical Structure

[0582] Fmoc-Val-Cit-PABC 10107 (prepared in Example 9, 0.30 g) was dissolved in DMF (5 mL), eribulin ERB (0.21 g) was added, DIPEA (0.32 mL) was added, and the reaction was carried out at room temperature for 2 hours. MTBE (30 mL) was added to obtain a precipitate, which was filtered and dried in vacuo to obtain 0.41 g of the product Fmoc-Val-Cit-PABC-ERB (10108-E).

[0583] Synthesis of Compound 10109-E

[0584]

Chemical Structure

[0585] Fmoc-Val-Cit-PABC-ERB (0.41 g) was dissolved in DMF (10 mL), and DEA (1 mL) was added. The solution turned purple-black, stirred at room temperature for 15 minutes, 100 mL of MTBE was added, filtered, and dried in vacuo to obtain 0.25 g of the white solid powder NH2-Val-Cit-PABC-ERB (10109-E).

[0586] Synthesis of Compound 101E00A

[0587]

Chemical Structure

[0588] 101B08 (0.10 g) was dissolved in 2 ml of DMF, 10109-E was added, PyBop (229 mg), DIPEA (0.10 ml) were added, and the mixture was stirred overnight at room temperature. The product was precipitated with methyl tert-butyl ether, the solid was dissolved in 30 ml of methanol, 10 ml of water was added, purified by ultrafiltration, and freeze-dried to obtain 110 mg of the final product (101E00A) as a white solid, HPLC purity: 98.0%.

[0589] Example 37: Synthesis of Compound 101P00A Synthesis of Compound 10108-P

[0590]

Chem.

[0591] Fmoc-Val-Cit-PABC (0.30 g) was dissolved in DMF (5 mL), parbocyclib PBC (0.15 g) was added, DIPEA (0.32 mL) was added, and the reaction was carried out at room temperature for 2 hours. MTBE (30 mL) was added to obtain a precipitate, which was filtered and dried under vacuum to obtain 0.35 g of the product Fmoc-Val-Cit-PABC-PAB (10108-P).

[0592] Synthesis of Compound 10109-P

[0593]

Chem.

[0594] Fmoc-Val-Cit-PABC-PAB (0.35 g) was dissolved in DMF (10 mL), and DEA (1 mL) was added. The solution turned purple-black, stirred at room temperature for 15 minutes, 50 mL of MTBE was added, filtered, and dried under vacuum to obtain 0.22 g of the white solid powder NH2-Val-Cit-PABC-PAB (10109-P).

[0595] Synthesis of Compound 101P00A

[0596]

Chem.

[0597] 101B08 (0.10 g) was dissolved in 2 ml of DMF, 10109-P was added, PyBop (200 mg) and DIPEA (0.10 ml) were added, and the mixture was stirred overnight at room temperature. The product was precipitated with methyl tert-butyl ether, the solid was dissolved in 30 ml of methanol, 10 ml of water was added, purified by ultrafiltration, and freeze-dried to obtain 135 mg of the final product (101P00A) as a white solid, HPLC purity: 98.5%.

[0598] Example 38: Synthesis of Compound 101P00B Compound 101P00B was synthesized according to the same method as in Examples 105 to 107:

[0599]

Chemical formula

[0600] Example 40: Synthesis of Compound 101P00C Compound 101P00C was synthesized according to the same method as in Examples 105 to 107:

[0601]

Chemical formula

[0602] Example 41: Synthesis of Compound 101P00D Compound 101P00D was synthesized according to the same method as in Examples 105 to 107:

[0603]

Chemical formula

[0604] Example 39: Synthesis of Compound 101P00E

[0605]

Chemical formula

[0606] To a solution of compound 101B08 (0.7 g) in DMF (6 mL), VC-PAB-MMAE (CAS: 644981-35-1) (0.54 g), PyBop (0.33 g) and DIPEA (0.18 g) were added at room temperature under nitrogen protection over 12 hours. MTBE was added to precipitate, and the precipitate was added to 60 ml of methanol and 70 ml of water, purified by ultrafiltration, and freeze-dried to obtain 0.8 g of the final product 101P00E, a pale yellow solid.

[0607] HPLC analysis method D, RT = 15.457 minutes.

[0608] Example 1: Synthesis of compound 101T19 Synthesis of compound 101T02

[0609]

Chemical formula

[0610] 32 g of hexanediamine was added to 120 ml of DCM, and a DCM solution of 15.60 g of 101T01 and 15 ml of triethylamine was slowly added via syringe, gradually changing to yellow, and reacting for 15 hours. The dot plate indicated that the reaction was complete. The precipitated solid was completely dissolved by adding DCM, and the organic phase was washed successively with water, saturated NaCl, saturated NaHCO3, and 2M NaOH, dried, and concentrated to obtain a pale yellow liquid.

[0611] 1 H NMR (400 MHz, DMSO-d6) δ 6.94 (t, J = 5.5 Hz, 1H), 4.16 ‐ 3.84 (m, 2H), 2.92 (td, J = 7.0, 5.8 Hz, 2H), 2.48 (t, J = 6.7 Hz, 2H), 1.68 ‐ 1.05 (m, 10H), 0.88 (s 9H). Synthesis of compound 101T03

[0612]

Chemical formula

[0613] 101B01 (8.541 g) was dissolved in 20 ml of DMSO / 10 ml of DMF. 1.692 g of 101T02 and 3.020 g of PyBop were added, 1.516 ml of DIPEA was added, protected with nitrogen, and stirred at room temperature for 16 hours. 6 ml of 0.5 M NaOH was added, the ACN precipitation product was added, pulverized, filtered, the filter cake was washed with acetonitrile, washed with water, and dried in vacuo to obtain 8.5 g of a white powder.

[0614] 1 H NMR (400 MHz, DMSO-d6) δ 7.32 (d, J = 2.2 Hz, 144H), 4.99 (s, 61H), 4.16 (br, 29H), 4.00 (t, J = 8.3 Hz, 2H), 3.85 (br, 30H), 3.22 ‐ 2.70 (m, 123H), 1.35 (s, 627H), 0.00 (s, 9H). Synthesis of Compound 101T04

[0615]

Chemical Structure

[0616] 8.5 g of 101T03 was dissolved in 85 ml of acetic acid, 28 ml of methanol was added, 1.7 g of 10% Pd / C was added, and hydrogenated at atmospheric pressure for 24 hours. The lower layer was padded with a 0.45 μm nylon filter membrane, the upper layer was compressed with diatomaceous earth, the reaction solution was pumped out, the diatomaceous earth was washed with methanol, the filtrate was concentrated, and dried in vacuo to obtain 8.57 g of a pale yellow solid (theoretical 7.23 g, 118%, structural unit 416.5). In the characteristic evaluation of NMR, 1.86 eq of HOAc (total amount 32.2 mmol, excess acetic acid 14.85 mmol) was obtained.

[0617] 1H NMR (400 MHz, Deuterium Oxide) δ 4.37 ‐ 4.09 (m, 31H), 3.97 (dd, J = 9.1, 5.3 Hz, 31H), 3.14 (s, 57H), 2.96 (t, J = 7.6 Hz, 55H), 1.98 (s, 168H), 1.86 ‐ 1.13 (m, 635H), 0.00 (s, 9H). Synthesis of Compound 101T18

[0618]

Chem.

[0619] 101T04 (3.79 g, 84.4%, 7.68 mmol) was dissolved in 15.4 ml of methanol (0.5 M).

[0620] 10104 - 2k (21.20 g) was dissolved in 20 ml of acetonitrile (0.5 M). Half of it was added to the 101T04 methanol solution, 2.676 ml of DIPEA was added, and the mixture was stirred at room temperature for 10 minutes. The remaining 10104 - 2k and 2.676 ml of DIPEA were added, and the mixture was stirred at room temperature for 4 hours. 175 ml of MTBE was gradually added to the reaction solution, and the mixture was left at 4 °C for 15 minutes. The upper layer liquid was discarded. The lower layer oil was added to 10 ml of acetonitrile, 150 ml of MTBE was added to precipitate the product, and the mixture was frozen at -20 °C until the product crystallized into a solid. The upper layer liquid was discarded, and the product was dried in vacuo to obtain an off-white powder. It was purified by ultrafiltration and freeze-dried to obtain 18.7 g of a white powder (5.5 mmol, 98.9%, structural unit 2364).

[0621] 1 H NMR (400 MHz, Deuterium Oxide) 4.17 (s, 37H), 4.02 (s, 98H), 3.66 (s, 5315H), 3.34 (s, 91H), 3.21 (s, 122H), 1.39 (s, 613H), 0.00 (s, 9H). Synthesis of Compound 101T19

[0622]

Chem.

[0623] 18.15 g of 101T18 was heated and dissolved in 80 ml of ethanol, 1.679 g of p-TsOH-H2O was added, dissolved by ultrasonic treatment, and the solvent was removed by spin distillation at 60 °C for 2 hours to concentrate. After cooling and vacuum drying, the product was obtained. 1 H NMR (400 MHz, Deuterium Oxide) δ 7.65 (d, J = 8.0 Hz, 59H), 7.33 (d, J = 7.9 Hz, 64H), 4.20 (d, J = 8.2 Hz, 43H), 4.02 (s, 87H), 3.67 (s, 5160H), 3.35 (s, 94H), 3.19 (t, J = 6.9 Hz, 134H), 2.36 (s, 96H), 2.03 ‐ 1.09 (m, 366H), 0.00 (s, 9H). Example 41: Synthesis of Compound 101H00A Synthesis of Compound 101H07

[0624]

Chem.

[0625] 30 g of 101H06, 21.69 g of Na2CO3, 300 mL of DMF were placed in a 300 mL single-necked flask, purged with nitrogen three times, and 29 g of CH3I was added by syringe and stirred at room temperature overnight. The reaction was completely controlled by LCMS. Methanol was removed by spin distillation, washed with 1N hydrochloric acid, and extracted with EtOAc. The organic phase was washed successively with 1N HCl and H2O, and the product was dried over anhydrous sodium sulfate and concentrated.

[0626] Synthesis of Compound 101H08

[0627]

Chem.

[0628] A 1 L three-port burner was pre-baked and evacuated to dryness, cooled to room temperature under nitrogen protection, 40 g of 101H07, 64.04 ml of DMP, 3.71 g of p-TsOH-H2O, and 500 ml of toluene were added. After three nitrogen replacements, the reaction was carried out under reflux at 130 °C overnight. The LCMS result showed a complete reaction, and toluene was removed by spin distillation. After chromatography on a DCM / EA column (PE:EA = 100:1 to 10:1), recrystallization was carried out with DCM / petroleum ether. HPLC 98.7%.

[0629] 1 H NMR (400 MHz, DMSO-d6) δ 9.86 (d, J = 7.1 Hz, 1H), 6.85 ‐ 6.69 (m, 2H), 6.63 (dd, J = 7.9, 1.7 Hz, 1H), 4.54 (ddd, J = 11.4, 7.0, 4.7 Hz, 1H), 3.67 (s, 3H), 3.09 (dd, J = 13.8, 4.9 Hz, 1H), 2.90 (dd, J = 13.9, 10.6 Hz, 1H), 1.60 (d, J = 5.8 Hz, 6H). Synthesis of Compound 101H09

[0630]

Chemical Structure

[0631] 15 g of 101H08 was dissolved in 60 mL of THF. After the solution became clear, 65 mL of 2M aqueous LiOH solution was added, and the mixture was stirred at room temperature for 2 hours. THF was removed by spin distillation, the pH was adjusted to 5 - 6 with 6N hydrochloric acid, and freeze-dried. The product was obtained by dissolving in EtOAc, filtering, and spin-drying.

[0632] Synthesis of Compound 101H17

[0633]

Chemical Structure

[0634] Dissolve 6 g of 101H09 in 50 ml of THF, add 25 ml of 2M aqueous sodium carbonate solution, add 6.62 g of anhydrous Boc to the above system in an ice bath, change the solution from clear to turbid, and stir overnight at room temperature.

[0635] As a result of TLC and LCMS, the control reaction was completed. Spin off THF, wash with 1N hydrochloric acid, and extract with EA. The organic phase was washed successively with 1N HCl and saturated NaCl, dried, and concentrated. Excess Boc anhydride was removed by DCM column chromatography, and the product was concentrated by spin distillation.

[0636] Synthesis of Compound 101H18

[0637]

Chemical formula

[0638] Add 3 g of 101H17 and 1.54 g of NHS to 20 ml of DCM, add 2.56 g of EDCI under an ice bath, remove the ice bath, and stir the reaction at room temperature for 4 hours. As a result of TLC and LCMS, the reaction was completed. Wash with 1N hydrochloric acid and saturated NaCl, dry with anhydrous sodium sulfate, perform PE / EA column chromatography (100:1 to 10:1), and spin dry to obtain a white solid.

[0639] Synthesis of Compound 101H19:

[0640]

Chemical formula

[0641] 101B07 (3.0 g, 1.26 mmol) was dissolved in 16 ml of DMF and added. 101H18 (1.729 g, 2.03 eq.) was added, 1.344 ml of DIPEA was added, and the mixture was stirred at room temperature for 18 hours. 75 ml of MTBE was added to the reaction solution, and it was centrifuged. The supernatant was discarded, and the viscous layer in the lower layer was washed once with MTBE. 3 ml of ACN was added, 40 ml of MTBE was added to precipitate, and it was dried under vacuum to obtain 3.4 g of a white solid.

[0642] 1 H NMR (400 MHz, Deuterium Oxide) δ 7.01 ‐ 6.48 (m, 93H), 4.32 (s, 33H), 4.20 (d, J = 7.4 Hz, 67H), 4.03 (s, 55H), 3.71 (s, 4631H), 3.39 (s, 90H), 3.24 (s, 415H), 1.23 (s, 1098H). Synthesis of compound 101H20:

[0643]

Chemical formula

[0644] 3.0 g of 101H19 was dissolved in 16 ml of DCM, 2.5% TIS, 0.6% water, and 25% TFA were added, and the mixture was stirred at room temperature for 16 hours. Most of the solvent was removed by spin distillation, MTBE was precipitated, ACN was dissolved, and the precipitation with MTBE was repeated twice, followed by drying under vacuum to obtain 2.463 g of product 101H20 (yield 83%), and the levodopa grafting rate = 112.48 / 3 / 31 = 120%.

[0645] 11H NMR (400 MHz, Deuterium Oxide) δ 6.94 ‐ 6.45 (m, 112H), 4.23 (s, 35H), 4.10 (d, J = 19.4 Hz, 62H), 3.94 (s, 53H), 3.62 (s, 4028H), 3.30 (s, 90H), 3.09 (s, 213H), 1.41 (t, J = 81.2 Hz, 351H). Synthesis of Compound 101H00A:

[0646]

Chem.

[0647] Weighed 201 mg of 101H20 and 30.0 mg of bortezomib into a 10 ml eggplant flask, added 2 ml of acetonitrile to dissolve, removed the solvent by spin distillation, added 3 ml of 20 mM pH 8.0 phosphate buffer, sonicated to dissolve the solid content, and freeze-dried to obtain 101H00A as a white freeze-dried block.

[0648] Example 42: Synthesis of Compound 101T00A-C4 Synthesis of Compound 10137C

[0649]

Chem.

[0650] Added 2 g of PTX and 0.4688 g of succinic anhydride to a 50 mL flask, evacuated with nitrogen, added 20 ml of anhydrous DCM and 1.632 ml of DIPEA to obtain a colorless transparent solution, and stirred the reaction at 25 °C for 4 hours. The reaction was completed by TLC (DCM / MeOH 15:1). Diluted with 20 mL of DCM, washed with 0.5 M hydrochloric acid (20 mL × 2), and spin-dried the organic phase to obtain a white solid. Pulverized with 40 mL of water for 20 minutes, filtered, washed, and freeze-dried to obtain 2.0909 g of the product (yield 93.6%).

[0651] 101T00A-C4

[0652]

Chem.

[0653] Dissolve 2.800 g of 101T19 in 15 ml of DMF, add 1.644 g of 10137C (purity 86.8%, 1.89 g), and cool in an ice bath. Add 0.897 g of PyBop, add 0.801 ml of DIPEA, and stir the reaction under nitrogen protection for 16 hours. Add 25 ml of MTBE precipitate in two portions, centrifuge, wash the low-viscosity liquid with 20 ml of 1:3 EtOAc / MTBE, and centrifuge. Dissolve in 10 ml of EtOAc, add 25 ml of MTBE, centrifuge, dry the precipitate under vacuum, heat and dissolve in 50 ml of methanol, filter through a 0.45 μm membrane, add 50 ml of methanol and 100 ml of water, and concentrate to 100 ml by ultrafiltration. Supplement with 50% methanol, concentrate to 900 ml after filtration, and lyophilize to obtain 2.784 g of a white solid (structural unit 3156, 0.88 mmol, 77%), and the theoretical coupling amount is 853.9 / 3156 = 27 wt.%.

[0654] 1 H NMR (400 MHz, Methanol-d4) δ 8.27 ‐ 8.00 (61H), 8.00 ‐ 7.10 (363H), 6.59 ‐ 6.37 (28H), 6.21 ‐ 5.99 (34H), 5.94 ‐ 5.78 (31H), 5.77 ‐ 5.37 (70H), 4.57 ‐ 4.06 (155H), 4.05 ‐ 3.90 (78H), 3.86 ‐ 3.49 (5160H), 3.38 ‐ 3.33 (91H), 3.27 ‐ 3.08 (79H), 2.96 ‐ 2.30 (255H), 2.28 ‐ 0.80 (857H). Example 43: Synthesis of Compound 101T00A-C5 Synthesis of Compound 10137

[0655] [Chemical formula]

[0656] 1.5 g of PTX and 0.3007 g of 101M22 were added to a 100 mL flask. After being evacuated and filled with nitrogen three times, 15 ml of anhydrous DCM and 0.918 ml of DIPEA were added to obtain a colorless transparent solution, which was reacted at room temperature for 5 hours. TLC (DCM / MeOH 10:1) indicated that the reaction was complete. 10 mL of DCM diluent was added, and it was washed with 1M hydrochloric acid (20 mL × 2). A white precipitate formed, which was filtered. The aqueous phase of the filtrate was discarded, and the organic phase was spin-dried to obtain a white solid. The cake was washed with water, combined with the above solid, pounded with 50 mL of water for 30 minutes, filtered, washed with water, and vacuum-dried to obtain 1.6542 g of the product, with a yield of 97.3% and a white solid powder. The HPLC purity was 99.61%.

[0657] Synthesis of Compound 101T00A-C5

[0658] [Chemical formula]

[0659] 1.80 g of 101T19 was dissolved in 9 ml of DMF, and 10137 (1.076 g, 99.6%) was added, using an ice bath. 0.577 g of PyBop was added, and 0.515 ml of DIPEA was added. The reaction was stirred under nitrogen protection for 15 hours.

[0660] The product was precipitated by adding 25 ml of MTBE, centrifuged, and the low-viscosity liquid was washed with 20 ml of 1:3 EtOAc / MTBE and centrifuged. It was dissolved in 10 ml of EtOAc, 25 ml of MTBE was added, centrifuged, and the precipitate was vacuum-dried.

[0661] The crude product was heated and dissolved in 25 ml of methanol, filtered through a 0.45 μm membrane, 25 ml of methanol and 50 ml of water were added, concentrated to 70 ml by ultrafiltration, supplemented with 50% methanol, filtered and concentrated to a volume of 700 ml, and freeze-dried to obtain 1.7794 g (structural unit 3170, 0.56 mmol, 76%) of a white solid. Theoretical coupling amount 853.9 / 3170 = 26.9 wt.%.

[0662] 1 H NMR (400 MHz, Methanol-d4) δ 8.27 ‐ 8.00 (65H), 8.00 ‐ 7.10 (395H), 6.60 ‐ 6.34 (34H), 6.20 ‐ 5.99 (34H), 5.95 ‐ 5.76 (34H), 5.74 ‐ 5.56 (33H), 5.56 ‐ 5.34 (37H), 5.13 ‐ 4.92 (38H), 4.53 ‐ 4.04 (158H), 4.04 ‐ 3.87 (68H), 3.86 ‐ 3.49 (5160H), 3.38 ‐ 3.33 (83H), 3.27 ‐ 2.93 (127H), 2.67 ‐ 0.84 (1206H). Example 44: Synthesis of Compound 101T00A-O: Synthesis of Compound 10137O

[0663]

Chemical Structure

[0664] 1.500 g of PTX and 0.3058 g of 101M24 were added to a 100 mL flask. After performing vacuum nitrogen filling three times, 15 ml of anhydrous DCM and 0.918 ml of DIPEA were added to obtain a colorless transparent solution. (Water bath at 10 °C). Stirred at room temperature for 6 hours. TLC (DCM / MeOH 10:1), the reaction was complete. 10 mL of DCM diluent was added, washed with 1M hydrochloric acid (20 mL × 2), white precipitate formed, filtered, the aqueous phase of the filtrate was discarded, and the organic phase was spin-dried to obtain a white solid; the filter cake was washed with water, combined with the above solid, and stored in the refrigerator (-20 °C). Pulverized with 50 mL of water for 30 minutes, filtered, washed, and vacuum dried to obtain 1.6386 g of the product, with a yield of 96.2%, a white solid powder.

[0665] Synthesis of compound 101T00A-O:

[0666]

Chemical formula

[0667] 1.80 g of 101T19 was dissolved in 9 ml of DMF, 10137O (92.6%, 1.156 g) was added, and it was placed in an ice bath. 0.577 g of PyBop was added, 0.515 ml of DIPEA was added, and it was stirred at room temperature for 16 hours under nitrogen protection.

[0668] The product was precipitated by adding 25 ml of MTBE, centrifuged, the low-viscosity liquid was washed with 20 ml of 1:3 EtOAc / MTBE, centrifuged, dissolved in 10 ml of EtOAc, 25 ml of MTBE was added, centrifuged, and the precipitate was vacuum dried.

[0669] The crude product was heated and dissolved in 25 ml of methanol, filtered through a 0.45 μm membrane, 25 ml of methanol and 50 ml of water were added, concentrated to 70 ml by ultrafiltration, supplemented with 50% methanol, filtered and concentrated in a 700 ml volume, and freeze-dried to obtain 1.7743 g (structural unit 3174, 0.56 mmol, 75.5%) of a white solid. 853.9 / 3174 = 26.9 wt.% 1 H NMR (400 MHz, Methanol-d4) δ 8.27 ‐ 8.00 (61H), 8.00 ‐ 7.10 (403H), 6.62 ‐ 6.35 (27H), 6.25 ‐ 6.00 (32H), 5.98 ‐ 5.81 (31H), 5.77 ‐ 5.42 (65H), 5.13 ‐ 4.93 (34H), 4.67 ‐ 4.00 (268H), 4.00 ‐ 3.87 (69H), 3.86 ‐ 3.50 (5160H), 3.38 ‐ 3.33 (82H), 3.27 ‐ 2.89 (117H), 2.71 ‐ 0.79 (1032H). Example 45: Synthesis of Compound 101T00A-S Synthesis of Compound 10137S

[0670]

Chemical Structure

[0671] 1.500 g of PTX and 0.6963 g of 101M25 were added to a 100 mL flask. After filling the flask with nitrogen three times under vacuum, 15 ml of anhydrous ACN and 1.836 ml of DIPEA were added, and the reaction was carried out at 41 °C for 5 hours in a water bath. By TLC (DCM / MeOH 10:1), the reaction was completed. ACN was removed by spin evaporation, 25 mL of DCM was added to dissolve the residue, and the solution was washed with 1M hydrochloric acid (20 mL × 2). The organic phase was spin-dried, triturated with 50 mL of water for 30 minutes, filtered, washed with water, and dried under vacuum. After column chromatography, the product was 1.58 g.

[0672] Synthesis of Compound 101T00A-S

[0673]

Chemical Structure

[0674] Dissolve 1.800 g of 101T19 in 9 ml of DMF, add 10137S (80%, 1.365 g), and cool in an ice bath. Add 0.577 g of PyBop, add 0.515 ml of DIPEA, and protect with nitrogen. Stir at room temperature for 16 hours. Add 25 ml of MTBE to precipitate the product, centrifuge, wash the low-viscosity liquid with 20 ml of 1:3 EtOAc / MTBE, centrifuge, add 10 ml of EtOAc to dissolve, add 25 ml of MTBE, centrifuge, and vacuum dry the precipitate to obtain 101T00A-S.

[0675] Heat and dissolve the crude 101T00A-S in 25 ml of methanol, filter through a 0.45 μm membrane, add 25 ml of methanol and 50 ml of water, emulsify, add 50 ml of methanol, concentrate to 70 ml by ultrafiltration, supplement this solution with 67% methanol, concentrate to 700 ml by filtration, and lyophilize to obtain 1.8641 g of a light brown solid (structural unit 3206, 0.581 mmol, 78.6%). The theoretical coupling amount of PTX 853.9 / 3174 = 26.6 wt.%.

[0676] 1 H NMR (400 MHz, Methanol-d4) δ 8.27 ‐ 8.00 (56H), 8.00 ‐ 7.10 (362H), 6.57 ‐ 6.33 (22H), 6.30 ‐ 6.00 (37H), 6.00 ‐ 5.79 (30H), 5.79 ‐ 5.41 (65H), 5.10 ‐ 4.94 (28H), 4.56 ‐ 4.03 (138H), 4.02 ‐ 3.86 (67H), 3.86 ‐ 3.49 (5160H), 3.38 ‐ 3.33 (103H), 3.26 ‐ 2.93 (139H), 2.68 ‐ 0.79 (1013H). Example 46: Synthesis of Compound 101T00A-NMe Synthesis of Compound 10137N

[0677]

Chemical Structure

[0678] 2.0087 g of PTX and 0.6075 g of 101M23 were added to a 100 mL three-necked flask, filled with vacuum nitrogen, 20 mL of anhydrous DCM and 1.639 mL of DIPEA were added, and the mixture was stirred and reacted at 25 °C for 5 hours. The reaction was completed by TLC (DCM / MeOH 10:1), washed three times with 0.5 M hydrochloric acid to precipitate a white solid. TLC (DCM / MeOH 10:1), the reaction was completed, washed three times with 0.5 M HCl to precipitate a white solid, washed with water, filtered, and dried in vacuo to obtain 2.27 g of the product.

[0679] Synthesis of Compound 101T00A-NMe

[0680]

Chemical formula

[0681] 1.400 g of 101T19 was dissolved in 7.5 ml of DMF, 10137N (93%, 0.911 g) was added, and the mixture was ice-bathed. 0.448 g of PyBop was added, 0.400 ml of DIPEA was added, and the mixture was stirred at room temperature for 16 hours under nitrogen protection.

[0682] The product was precipitated with 25 ml of MTBE, centrifuged, the low-viscosity liquid was washed with 20 ml of 1:3 EtOAc / MTBE, centrifuged, dissolved in 10 ml of EtOAc, 25 ml of MTBE was added, centrifuged, and the precipitate was dried in vacuo.

[0683] The crude product was heated and dissolved in 25 ml of methanol, filtered through a 0.45 μm membrane, 25 ml of methanol and 50 ml of water were added, concentrated to 70 ml by ultrafiltration, supplemented with 50% methanol, filtered and concentrated in a 700 ml volume, and freeze-dried to obtain 1.4407 g (structural unit 3187, 0.452 mmol, 79.3%) of a white solid. The theoretical coupling amount 853.9 / 3187 = 26.9 wt.%.

[0684] 11H NMR (400 MHz, Methanol-d4) δ 8.27 ‐ 8.00 (60H), 7.99 ‐ 7.10 (379H), 6.60 ‐ 6.35 (30H), 6.25 ‐ 5.99 (33H), 5.99 ‐ 5.81 (31H), 5.76 ‐ 5.52 (56H), 5.19 ‐ 4.93 (36H), 4.56 ‐ 4.04 (154H), 4.04 ‐ 3.87 (71H), 3.86 ‐ 3.49 (5160H), 3.38 ‐ 3.33 (82H), 3.27 ‐ 2.94 (155H), 2.64 ‐ 0.82 (1073H). Example 50: Determination of the PTX Coupling Amount Composition of the Polymer Solution: Weighed 101T00A-C4, 101T00A-C5, 101T00A-O, 101T00A-S, 101T00A-NMe into a volumetric flask, fixed the ACN to 10.00 ml, weighed the mass of the added ACN, averaged the density of ACN from the masses of the six flasks, and took the density as 0.7765 g / cm 3 and calculated the added amount of ACN from the mass of the solution.

[0685] [Table 7]

[0686] [Table 8]

[0687] The PTX standard curve by HPLC was y = 8.776x + 485.47, R 2 = 0.9984.

[0688] The PTX coupling amount was calculated by HPLC:

[0689] [Table 9]

[0690] 10 - 15 mg of the polymer was dissolved in CD3OD, and PEG was calculated with 44 repeating units. Since the same batch of raw material 101T19 was used, the PEG integral value was fixed at the integral value of 101T19, which is 5159.96. The range of the integral of the PEG repeating unit -CH2CH2O- was fixed at 3.49 - 3.86, and the number of single-stranded polymer PEG chain linkages: PEG 3.49 - 3.86 integral / (44 * 4) = 29.

[0691] The number of hydrogens on the benzene ring of PTX is 15, and among them, the hydrogen integral from 7.10 to 8.00 ppm should be 13. The number of couplings of single-stranded polymer PTX: PTX 7.10 - 8.00 integral value / 13.

[0692]

Number

[0693] The amount of PTX coupling was calculated by NMR:

[0694]

Table 10

[0695] Example 51: Synthesis of Compound 101N19 Synthesis of Compound 101N01

[0696]

Chemistry

[0697] 101N-SM (3.2 g, 0.01 mol), NHS (1.36 g, 0.011 mol), DCC (2.43 g, 0.011 mmol) and DMF (60 mL) were added to a 100 mL single-necked vial, stirred at room temperature for 16 hours, sampled for LCMS detection. After the raw material reaction was completed, the filtration cake was washed with dichloromethane and concentrated under reduced pressure to obtain a crude product. The crude product was dissolved in 20% isopropanol / ethyl acetate, triturated for 2 hours, filtered, the filtration cake was washed with ethyl acetate, and the filtration cake was scooped out and dried. The crude product was dissolved in 20% isopropanol / ethyl acetate, triturated for 2 hours, filtered, the filtration cake was washed with ethyl acetate, and the filtration cake was scooped out and dried to obtain 3.89 g of a white solid, yield: 92.6%, purity: 95.6%.

[0698] Synthesis of Compound 101N02

[0699]

Chemical Structure

[0700] 101N01 (1 g, 2.53 mmol), L-glutamic acid-5-tert-butyl ester (565 mg, 2.78 mmol), DIPEA (0.98 g, 7.58 mmol) and DMF (45 mL) were added to a 100 mL single-necked flask. Nitrogen replacement was carried out 3 times to protect. Stir at room temperature for 16 hours, sample for LCMS. After the raw material reaction was completed, the reaction solution was directly concentrated to obtain 3 g of a crude product. 1N hydrochloric acid (50 mL) was added and triturated for 30 minutes, filtered, the filtration cake was washed with water and then with ethyl ether, and the solid was dried to obtain 1.07 g of a white solid, yield: 87.7%. The LCMS was correct, HPLC: 91.2%.

[0701] 11H NMR (400 MHz, DMSO-d6) δ 11.25 (s, 1H), 8.52 (d, J = 7.7 Hz, 1H), 7.80 (d, J = 8.1 Hz, 1H), 7.29 (d, J = 8.1 Hz, 1H), 6.46 (d, J = 2.2 Hz, 1H), 4.39 (ddd, J = 9.9, 7.7, 4.8 Hz, 2H), 3.12 ‐ 2.83 (m, 2H), 2.33 (t, J = 7.6 Hz, 1H), 2.17 ‐ 1.85 (m, 1H), 1.39 (s, 3H). Synthesis of Compound 101N03

[0702] [Chemical formula]

[0703] 101T19 (1 g, 0.41 mmol), 101N02 (298 mg, 0.62 mmol) and DMAC (15 mL) were added to a 10 mL single vial, dissolved by ultrasound, cooled to 0 °C with ice water, PyBOP (321 mg, 0.62 mmol) and DIPEA (213 mg, 1.64 mmol) were added sequentially, replaced three times under nitrogen gas protection, stirred at 0 °C for 10 minutes, naturally warmed to room temperature and stirred for 16 hours, sampled for HPLC detection, the reaction of the raw materials was completed, MTBE (80 mL) for reaction precipitation at -68 °C, centrifuged, the supernatant was poured out, the solid and 1, 2 were combined by extraction and drying to obtain 1.21 g of a yellow solid. The crude product was dissolved in a 50 mL (methanol:water = 1:1) methanol-water mixed solution, ultrafiltered with 480 mL (methanol:water = 1:1), the filtrate was lyophilized to obtain 954 mg of a pale yellow solid, purity: 99.32%.

[0704] 11H NMR (400 MHz, Methanol-d4) δ 7.71 (d, J = 26.5 Hz, 60H), 7.24 (s, 62H), 6.30 (s, 27H), 4.68 ‐ 4.05 (m, 158H), 4.05 ‐ 3.85 (m, 92H), 3.84 ‐ 3.37 (m, 7128H), 3.37 ‐ 3.25 (m, 334H), 3.23 ‐ 2.59 (m, 226H), 2.57 ‐ 2.27 (m, 73H), 2.25 ‐ 0.90 (m, 872H), 0.00 (s, 9H). Synthesis of Compound 101N19

[0705]

Chemical Structure

[0706] 101N03 (954 mg, 0.011 mmol), trifluoroacetic acid (3 mL) and DCM (12 mL) were added to a 50 mL single-necked vial, stirred at room temperature for 2 hours, the raw material for HPLC detection was sampled, the reaction solution was concentrated and directly dried to obtain a crude product. MTBE (10 mL) was added to the crude product, ultrasonic treatment was carried out, the supernatant was poured out, and the solid was vacuum dried to obtain a brown solid. Solid 901 mg, purity: 98.55%. The correct product was confirmed by HNMR.

[0707] 1 1H NMR (400 MHz, Methanol-d4) δ 8.04 ‐ 7.56 (m, 60H), 7.49 ‐ 7.05 (m, 61H), 6.69 ‐ 6.26 (m, 23H), 4.67 ‐ 4.12 (m, 111H), 4.11 ‐ 3.87 (m, 66H), 3.86 ‐ 3.41 (m, 4963H), 3.40 ‐ 3.29 (m, 492H), 3.27 ‐ 3.13 (m, 318H), 3.10 ‐ 2.83 (m, 119H), 2.66 ‐ 2.40 (m, 72H), 2.35 ‐ 2.07 (m, 69H), 1.96 ‐ 1.26 (m, 344H). Example 47: Synthesis of Compound 101N20 Synthesis of Compound 101N04

[0708]

Chem.

[0709] 101N01 (760 mg, 1.9 mmol), 1-tert-butyl-L-glutamic acid (586 mg, 2.88 mmol), DIPEA (745 mg, 5.7 mmol) and DMF (40 mL) were added to a 50 mL single neck vial. The vial was purged with nitrogen three times, protected, stirred at room temperature for 16 hours, and LCMS was sampled. When the reaction of the starting material was completed, the pH was adjusted to 4 with 1N hydrochloric acid, and extraction with dichloromethane was carried out three times. The combined organic phases were washed once with saturated brine, and water was sent to the organic phase for LCMS detection. The product in the aqueous phase was concentrated under reduced pressure to obtain 1.11 g of crude product.

[0710] 1 H NMR (400 MHz, DMSO-d6) δ 12.18 (s, 1H), 10.61 (d, J = 2.2 Hz, 1H), 10.15 (s, 1H), 8.54 (d, J = 7.5 Hz, 1H), 7.78 (d, J = 8.1 Hz, 2H), 7.29 (d, J = 8.0 Hz, 2H), 6.31 (d, J = 2.1 Hz, 1H), 6.00 (s, 2H), 4.32 (ddd, J = 9.6, 7.4, 5.1 Hz, 1H), 2.98 (dd, J = 9.4, 6.1 Hz, 2H), 2.86 (dd, J = 9.4, 6.1 Hz, 2H), 2.36 (t, J = 7.5 Hz, 2H), 2.12 ‐ 1.88 (m, 2H), 1.41 (s, 9H). Synthesis of Compound 101N05

[0711]

Chem.

[0712] 101T19 (1 gram, 0.41 mmol), 101N04 (298 mg, 0.62 mmol) and DMAC (15 mL) were added to a 10 mL vial, dissolved by ultrasound, cooled to 0 °C with ice water, and PyBOP (321 mg, 0.62 mmol) and DIPEA (213 mg, 1.64 mmol) were sequentially added, followed by nitrogen substitution. Protected three times, stirred at 0 °C for 10 minutes, and naturally warmed to room temperature with stirring for 16 h. Sampling was performed for HPLC detection. After the reaction of the raw materials was completed, MTBE (80 mL) for reaction precipitation was added at -68 °C, followed by centrifugation. The supernatant was poured out, and the crude product was dissolved in 50 mL of a methanol-water mixed solution (methanol:water = 1:1) to obtain 1.21 g of a gray solid by solid pull drying. The crude product was dissolved in 480 mL of a methanol-water mixed solution (methanol:water = 1:1) for ultrafiltration, and the filtrate was freeze-dried to obtain 951 mg of a pale yellow solid, purity: 99.86%.

[0713] 1 H NMR (400 MHz, Methanol-d4) δ 7.72 (s, 60H), 7.24 (s, 62H), 6.32 (s, 36H), 4.65 ‐ 4.13 (m, 153H), 3.90 (s, 64H), 3.77 (t, J = 4.9 Hz, 48H), 3.59 (s, 5210H) 3.37 ‐ 3.23 (m, 345H), 3.12 (d, J = 16.7 Hz, 89H), 2.96 (s, 131H), 2.41 (s, 63H), 2.01 (s, 41H), 2.17 (dd, J = 17.4, 10.0 Hz, 43H), 1.64 (d, J = 58.7 Hz, 128H), 1.41 (s, 540H), 0.00 (s, 9H). Synthesis of Compound 101N20

[0714]

Chemical Structure

[0715] 101N05 (951 mg, 0.011 mmol), trifluoroacetic acid (3 mL), and DCM (12 mL) were added to a 50 mL single-necked vial, stirred at room temperature for 2 hours, the raw material for HPLC detection was sampled, the reaction solution was concentrated, directly dried to obtain a crude product, MTBE (10 mL) was added to the crude product, ultrasonic treatment was performed, the supernatant was poured out, and the solid was dried in vacuo to obtain a brown solid, 820 mg, purity: 99%. The correct product was confirmed by HNMR.

[0716] 1 H NMR (400 MHz, Methanol-d4) δ 7.80 (s, 60H), 7.30 (s, 60H), 6.46 (s, 27H), 4.62 (s, 32H), 4.33 (d, J = 26.9 Hz, 53H), 3.97 (s, 57H), 3.64 (d, J = 4.7 Hz, 4032H), 3.37 (s, 62H), 3.33 (p, J = 1.7 Hz, 298H), 3.27 ‐ 2.83 (m, 185H), 2.50 (s, 58H), 2.27 (d, J = 44.3 Hz, 73H), 1.95 ‐ 1.01 (m, 336H). Example 48: Synthesis of Compound 101J00A-O: Synthesis of Compound 10133

[0717]

Chemical formula

[0718] 41.46 g of (Boc)2O was added to a DMF solution of 5.00 g of 101M20 and reacted at 37 °C for 18 hours (the water bath was set at 40 °C). The reaction was complete. MS gave a peak [M-tBu+H] = 364. 400 mL of water and 200 mL of PE / EA (1:1) were added to precipitate a white solid, which was filtered, and the filter cake was washed successively with water and PE / EA (1:1) and dried under vacuum to obtain a white solid powder product of 6.3160 g, yield 91.5%.

[0719] 11H NMR (400 MHz, DMSO) δ 10.50 (s, 1H), 8.18 (d, J = 7.6 Hz, 1H), 7.07 (d, J = 7.6 Hz, 1H), 6.35 ‐ 6.24 (m, 1H), 6.16 (t, J = 7.5 Hz, 1H), 5.29 (t, J = 5.5 Hz, 1H), 4.18 (tt, J = 12.8, 6.9 Hz, 1H), 3.88 (dt, J = 8.5, 3.0 Hz, 1H), 3.85 ‐ 3.60 (m, 2H), 1.46 (s, 9H). Synthesis of Compound 10134

[0720]

Chem.

[0721] Add 9.077 g of sodium carbonate to a 500 mL flask, add 62 ml of water and dissolve it. Add 6.2227 g of 10133 with stirring. Dissolve 3.738 g of (Boc)2O in 249 ml of dioxane and add it to the above system. A white precipitate was formed. React at 30 °C in a water bath for 48 hours. Add 250 mL of water, extract the precipitate with ethyl acetate (250 mL, 150 mL×2). Wash the organic phase with saturated NaCl (100 mL), dry over anhydrous sodium sulfate, spin dry, and vacuum dry. Column chromatography (silica gel 200 - 300 mesh, 300 mL). Wash the coupling 3 Boc impurities with PE:EA 2:1→1:1 and replace with DCM:MeOH 20:1. Recover the product, spin dry, and obtain 6.250 g of a white solid powder in a yield of 78.7%.

[0722] 11H NMR (400 MHz, DMSO) δ 10.57 (s, 1H), 8.11 (d, J = 7.7 Hz, 1H), 7.10 (d, J = 7.7 Hz, 1H), 6.28 (t, J = 8.7 Hz, 1H), 5.32 (dd, J = 6.4, 5.2 Hz, 1H), 5.28 ‐ 5.13 (m, 1H), 4.24 (dt, J = 7.0, 3.4 Hz, 1H), 3.79 (ddd, J = 12.7, 5.3, 3.2 Hz, 1H), 3.68 (ddd, J = 12.7, 6.5, 3.9 Hz, 1H), 1.47 (d, J = 2.9 Hz, 18H). Synthesis of Compound 10135O

[0723]

Chem.

[0724] 1.100 g of 10134 and 0.5510 g of 101M24 were added to a 50 mL flask, filled with nitrogen under vacuum, 11 ml of anhydrous DCM and 1.654 ml of DIPEA were added, and the reaction was stirred at 25 °C for 5 hours (the water bath was set at 30 °C). It was washed three times with 0.5 M hydrochloric acid, washed with water, and dried under vacuum to obtain 1.3342 g of a white powder, and the yield was 97.0%.

[0725] Synthesis of Compound 101J11O

[0726]

Chem.

[0727] 150 mg of 101T19 was dissolved in DMF, 53.5 mg of 10135O was added, and the mixture was placed in an ice bath. After 20 minutes, 48 mg of PyBop was added, 43 μl of DIPEA was added, and the mixture was protected with nitrogen. The reaction was stirred at room temperature for 18 hours. MTBE was added to precipitate the product, which was centrifuged and the precipitate was dried under vacuum. The crude product was dissolved in 5 ml of methanol, filtered through a 0.45 μm membrane, 5 ml of water was added, and the product was purified by ultrafiltration and lyophilized to obtain 138 mg of a white solid.

[0728] Synthesis of Compound 101J00A-O

[0729]

Chemical Structure

[0730] 101J11O (138 mg), trifluoroacetic acid (3 mL) and DCM (7 mL) were added, and the reaction solution was stirred at room temperature for 8 hours. The reaction solution was concentrated, MTBE was added to precipitate the product, which was centrifuged and dried under vacuum to obtain 101J00A-O as a white solid.

[0731] Example 49: Synthesis of Compound 101T28 Synthesis of Compound 101L09

[0732]

Chemical Structure

[0733] 0.470 g of 101L07 and 0.2988 g of 101L08 were dissolved in 4.7 ml of pyridine and stirred at 40 °C under nitrogen protection. The solution gradually changed to green, and after 1 hour, the product was precipitated with methyl tert-butyl ether, triturated with acetone, and dried under vacuum to obtain 0.661 g of a dark green powder product in a yield of 99.4% (Mw = 768, pyridine salt). HPLC purity was 87.47%.

[0734] Synthesis of Compound 101L10

[0735]

Chem.

[0736] 0.662 g of 101L09 was suspended in 12 ml of DCM, 0.246 g of 2-mercapto-thiazoline and 5.9 mg of DMAP were added, it was placed in an ice bath, 686 mg of EDCl was added, taken out from the ice bath, and stirred at room temperature for 2 hours. The reaction was completed. Precipitated with MTBE, dissolved in 6 mL of DCM, precipitated with 90 mL of acetone, and centrifuged. Beaten with 5 mL of acetone for 40 minutes and centrifuged. The product was dried in vacuo to give 0.7200 grams, yield 95%.

[0737] Synthesis of Compound 101T28

[0738]

Chem.

[0739] 200.0 mg of 101T19 was dissolved in 1 ml of DMF, 149 mg of 101L10 was added, 58 μl of DIPEA was added, the reaction was protected with nitrogen, and shielded from light at room temperature for 2 hours. 101L10 was consumed.

[0740] The product was precipitated in 20 mL of MTBE and centrifuged. The lower layer was a dark green oil, dissolved in 1 mL of MeOH, precipitated with 15 mL of MTBE, centrifuged, and the precipitate was dried in vacuo to obtain 190 mg of a dark green solid powder, yield 95%, HPLC retention time 9.698 minutes, and the maximum absorption wavelength of the compound in the 300 - 900 nm band was 786 nm.

[0741] Example 50: Synthesis of Compound 216A13

[0742]

Chem.

[0743] Synthesis of Compound 216A03: Under nitrogen protection at room temperature (25 °C), abiraterone (399 mg, 2.0 mmol, 1.0 eq), tert-butyl carboxylic acid (383 mg, 2.2 mmol, 1.1 eq), DMAP (293 mg, 2.4 mmol, 1.2 eq), EDC HCl (575 mg, 3.0 mmol, 1.5 eq), and anhydrous DCM (40 mL) were sequentially added. The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reactants was monitored by TLC. The solvent was spin-dried and separated by silica gel column chromatography (33 - 50% EtOAc in PE) to obtain the product 216A03, 1.1 g, white solid, in 99% yield.

[0744] 1 1H NMR (400 MHz, CDCl3) δ 8.61 (d, J = 2.2 Hz, 1H), 8.45 (d, J = 4.6 Hz, 1H), 7.68 ‐ 7.58 (m, 1H), 7.21 (dd, J = 8.0, 4.8 Hz, 1H), 5.99 (s, 1H), 5.40 (d, J = 5.1 Hz, 1H), 4.72 ‐ 4.56 (m, 1H), 2.64 ‐ 2.48 (m, 4H), 2.43 ‐ 2.19 (m, 3H), 2.09 ‐ 2.02 (m, 3H), 1.91 ‐ 1.82 (m, 2H), 1.75 ‐ 1.57 (m, 5H), 1.54 ‐ 1.42 (m, 10H), 1.27 ‐ 1.23 (m, 1H), 1.20 ‐ 1.06 (m, 5H), 1.04 (s, 3H). Synthesis of compound 216A01: TFA (5 mL) was added to a DCM solution (25 mL) of 216A03 (1.144 g, 2.26 mmol, 1.0 eq) at room temperature (25 °C). The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reactants was monitored by TLC. The solvent was spin-dried and separated by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain the product 216A01, 1.0 g, white solid, in 99% yield.

[0745] 11H NMR (400 MHz, CDCl3) δ 8.88 (s, 1H), 8.68 (d, J = 5.5 Hz, 1H), 8.24 (d, J = 8.2 Hz, 1H), 7.74 (dd, J = 8.1, 5.5 Hz, 1H), 6.31 (s, 1H), 5.41 (d, J = 5.0 Hz, 1H), 4.71 ‐ 4.57 (m, 1H), 2.74 ‐ 2.57 (m, 4H), 2.41 ‐ 2.23 (m, 3H), 2.17 ‐ 2.01 (m, 3H), 1.92 ‐ 1.43 (m, 10H), 1.15 ‐ 1.07 (m, 7H). Synthesis of Compound 216A13: Under nitrogen protection at room temperature (25 °C), 101T19 (200 mg, 0.082 mmol, 1.0 eq, based on the molar amount of the polymerization unit), 216A01 (55.4 mg, 0.12 mmol, 1.5 eq), PyBOP (64.0 mg, 0.12 mmol, 1.5 eq), DIPEA (0.09 mL, 0.49 mmol, 4.0 eq) and DMF (1.6 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reactants was monitored by HPLC. When MTBE (10 mL) was added to DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was washed successively with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low molecular weight impurities were removed by ultrafiltration with a MeOH / aqueous solution. By HPLC monitoring, the low molecular weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain 197 mg of Compound 216A13 as a white solid with a purity of 99%.

[0746] Example 51: Synthesis of Compound 216A14:

[0747] [Chemical formula]

[0748] Synthesis of Compound 216A08: Under nitrogen protection at room temperature (25 °C), abiraterone (2.0 g, 5.7 mmol, 1.0 eq), chloroacetyl chloride (1.73 g, 8.6 mmol, 1.5 eq), DIPEA (1.42 mL, 8.6 mmol, 1.5 eq) and anhydrous DCM (40 mL) were added sequentially. The reaction mixture was stirred at room temperature for 2 h, and the complete conversion of the reaction was monitored by HPLC. The reaction solution was used directly in the next step.

[0749] Synthesis of compound 216A09: Amine (2.14 mL, 11.4 mmol, 2.0 eq) was added to the above reaction solution, and the mixture was stirred at room temperature for 15 h. The solvent was spin-dried and separated by silica gel column chromatography (PE / EA = 3 / 1) to obtain the product 216A09, 2.83 g, white solid, 88% overall yield in two steps.

[0750] 1 H NMR (400 MHz, CDCl3) δ 8.61 (d, J = 2.2 Hz, 1H), 8.45 (dd, J = 4.8, 1.6 Hz, 1H), 7.64 (dt, J = 7.9, 2.0 Hz, 1H), 7.21 (dd, J = 7.9, 4.8 Hz, 1H), 6.04 ‐ 5.92 (m, 1H), 5.41 (d, J = 4.7 Hz, 1H), 4.60 ‐ 4.40 (m, 1H), 3.46 ‐ 3.23 (m, 4H), 2.98 ‐ 2.82 (m, 6H), 2.45 ‐ 2.21 (m, 3H), 2.11 ‐ 1.97 (m, 3H), 1.88 ‐ 1.55 (m, 9H), 1.45 (s, 9H), 1.20 ‐ 1.01 (m, 8H). Synthesis of compound 216A10: At room temperature (25 °C), TFA (5 mL) was added to a DCM solution (50 mL) of 216A09 (2.83 g, 5.0 mmol, 1.0 eq). The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reaction was monitored by HPLC. The solution was spin-dried to obtain a white solid.

[0751] Synthesis of Compound 216A11: Under nitrogen protection at room temperature (25 °C), 10107 (4.6 g, 5.0 mmol, 1.5 eq) and DIPEA (1.8 mL, 10.0 mmol, 2.0 eq) were added to a DMF solution (50 mL) of 216A10. The reaction mixture was stirred at room temperature for 15 hours, and the complete conversion of the reactants was monitored by HPLC. The reaction solution was used as it was without further treatment and directly proceeded to the next step.

[0752] Synthesis of Compound 216A12: Under nitrogen protection at room temperature (25 °C), diethylamine (2.6 mL, 25.0 mmol, 5.0 eq) was added to the above DMF solution. The reaction mixture was stirred at room temperature for 15 hours, and the complete conversion of the reactants was monitored by HPLC. The solvent was spin-dried and separated by silica gel column chromatography (5% - 25% MeOH in DCM) to obtain the product 216A12, 2.43 g, as a yellow solid. The overall yield in three steps was 56%.

[0753] 1 H NMR (400 MHz, CDCl3) δ 9.82 ‐ 9.58 (br, 1H), 8.59 (d, J = 2.2 Hz, 1H), 8.49 ‐ 8.32 (m, 2H), 7.63 (d, J = 8.0 Hz, 1H), 7.59 ‐ 7.46 (m, 2H), 7.24 ‐ 7.16 (m, 3H), 6.12 ‐ 5.87 (m, 2H), 5.41 ‐ 5.29 (m, 2H), 5.01 (br, 2H), 4.71 (br, 1H), 4.45 (br, 1H), 3.35 (br, 6H), 2.94 ‐ 2.70 (m, 7H), 2.48 ‐ 2.15 (m, 3H), 2.15 ‐ 1.92 (m, 4H), 1.88 ‐ 1.43 (m, 12H), 1.15 ‐ 0.96 (m, 8H), 0.85 (dd, J = 21.5, 6.6 Hz, 7H). Synthesis of Compound 216A14: Under nitrogen protection at room temperature (25 °C), 101B13 (100 mg, 0.04 mmol, 1.0 eq, based on the molar amount of the polymerization unit), 216A12 (52.1 mg, 0.06 mmol, 1.5 eq), PyBOP (31.2 mg, 0.06 mmol, 1.5 eq), DIPEA (0.03 mL, 0.16 mmol, 4.0 eq) and DMF (0.8 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reaction was monitored by HPLC. When MTBE (10 mL) was added to the DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was successively washed with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low-molecular-weight impurities were removed by ultrafiltration with an MeOH / aqueous solution. By HPLC monitoring, the low-molecular-weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain 117 mg of compound 216A14 as a white solid with a purity of 97.55%.

[0754] Example 52: Synthesis of Compound 216A15

[0755]

Chemical formula

[0756] Synthesis of Compound 216A07: Under nitrogen protection at room temperature (25 °C), abiraterone (1.0 g, 2.86 mmol, 1.0 eq), acid anhydride (1.11 g, 8.6 mmol, 3.0 eq), DMAP (697 mg, 5.7 mmol, 2.0 eq), DIPEA (1.0 mL, 5.7 mmol, 2.0 eq) and anhydrous DCM (10 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h, and the remaining starting materials were monitored by TLC. The solvent was spin-dried and separated by silica gel column chromatography (10 - 30% MeOH in DCM) to obtain 1.08 g of product 216A07 as a pale yellow solid in a yield of 79%.

[0757] 11H NMR (400 MHz, CDCl3) δ 8.61 (d, J = 2.2 Hz, 1H), 8.45 (dd, J = 4.8, 1.6 Hz, 1H), 7.69 ‐ 7.61 (m, 1H), 7.23 (dd, J = 7.9, 4.8 Hz, 1H), 6.02 ‐ 5.96 (m, 1H), 5.42 (d, J = 4.9 Hz, 1H), 4.76 ‐ 4.63 (m, 1H), 3.57 ‐ 3.34 (m, 4H), 2.57 (s, 3H), 2.39 ‐ 2.20 (m, 3H), 2.11 ‐ 1.99 (m, 3H), 1.93 ‐ 1.81 (m, 2H), 1.74 ‐ 1.56 (m, 5H), 1.50 ‐ 1.45 (m, 3H), 1.16 ‐ 1.06 (m, 4H), 1.04 (s, 3H). Synthesis of Compound 216A15: Under nitrogen protection at room temperature (25 °C), 101T19 (100 mg, 0.041 mmol, 1.0 eq, based on the molar amount of the polymerization unit), 216A07 (29.2 mg, 0.061 mmol, 1.5 eq), PyBOP (32.0 mg, 0.061 mmol, 1.5 eq), DIPEA (0.03 mL, 0.16 mmol, 4.0 eq) and DMF (0.8 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 hours, and the complete conversion of the reactants was monitored by HPLC. When MTBE (10 mL) was added to DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was continuously washed with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low-molecular-weight impurities were removed by ultrafiltration with a MeOH / aqueous solution. By HPLC monitoring, the low-molecular-weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain the product 216A15, 92 mg, white solid, purity 99.13%.

[0758] Example 53: Synthesis of Compound 216A16:

[0759] [Chemical formula]

[0760] Synthesis of Compound 216A02: Under nitrogen protection at room temperature (25 °C), abiraterone (350 mg, 1.0 mmol, 1.0 eq), acid anhydride (342 mg, 3.0 mmol, 3.0 eq), DMAP (244 mg, 2.0 mmol, 2.0 eq), DIPEA (0.35 mL, 2.0 mmol, 2.0 eq) and anhydrous DCM (50 mL) were added sequentially. The reaction mixture was stirred at 40 °C for 15 h. The remaining starting materials in the reaction were monitored by TLC. The solvent was spin-dried and separated by silica gel column chromatography (5 - 20% MeOH in DCM) to obtain the product 216A02, 382 mg, white solid, in 82% yield.

[0761] 1 H NMR (400 MHz, CDCl3) δ 8.62 (d, J = 2.2 Hz, 1H), 8.46 (d, J = 4.7 Hz, 1H), 7.68 (d, J = 7.9 Hz, 1H), 7.26 ‐ 7.23 (m, 1H), 6.13 ‐ 5.93 (m, 1H), 5.41 (d, J = 5.1 Hz, 1H), 4.70 ‐ 4.53 (m, 1H), 2.46 ‐ 2.26 (m, 10H), 2.08 ‐ 1.95 (m, 5H), 1.90 ‐ 1.83 (m, 2H), 1.71 ‐ 1.52 (m, 5H), 1.17 ‐ 1.07 (m, 4H), 1.04 (s, 3H). Synthesis of Compound 216A16: Under nitrogen protection at room temperature (25 °C), 101T19 (200 mg, 0.082 mmol, 1.0 eq, based on the molar amount of the polymerization unit), 216A02 (57.0 mg, 0.12 mmol, 1.5 eq), PyBOP (64.0 mg, 0.12 mmol, 1.5 eq), DIPEA (0.09 mL, 0.49 mmol, 4.0 eq) and DMF (1.6 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 hours, and the complete conversion of the reactants was monitored by HPLC. When MTBE (10 mL) was added to the DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was washed successively with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low-molecular-weight impurities were removed by ultrafiltration with an MeOH / aqueous solution. By HPLC monitoring, the low-molecular-weight impurities were completely removed. The MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain the product 216A16, 236 mg, white solid, purity 99.60%.

[0762] Example 54: Synthesis of Compound 216A17

[0763] [Chemical formula]

[0764] Synthesis of Compound 216A05: Under nitrogen protection at room temperature (25 °C), abiraterone (1.0 g, 2.86 mmol, 1.0 eq), acid anhydride (1.0 g, 8.6 mmol, 3.0 eq), DMAP (697 mg, 5.7 mmol, 2.0 eq), DIPEA (1.0 mL, 5.7 mmol, 2.0 eq) and anhydrous DCM (10 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 hours, and the remaining starting materials were monitored by TLC. The solvent was spin-dried and separated by silica gel column chromatography (5 - 20% MeOH in DCM) to obtain the product 216A05, 934 mg, white solid, yield 70%.

[0765] 11H NMR (400 MHz, CDCl3) δ 8.65 (d, J = 2.3 Hz, 1H), 8.47 (d, J = 4.9 Hz, 1H), 7.75 (dt, J = 7.9, 1.9 Hz, 1H), 7.32 (dd, J = 8.0, 4.9 Hz, 1H), 6.10 ‐ 5.99 (m, 1H), 5.42 (d, J = 5.0 Hz, 1H), 4.79 ‐ 4.68 (m, 1H), 4.25 (s, 4H), 2.44 ‐ 2.23 (m, 3H), 2.14 ‐ 1.99 (m, 3H), 1.96 ‐ 1.83 (m, 2H), 1.79 ‐ 1.43 (m, 8H), 1.16 ‐ 1.06 (m, 4H), 1.04 (s, 3H). Synthesis of Compound 216A17 Under nitrogen protection at room temperature (25 °C), 101T19 (200 mg, 0.082 mmol, 1.0 eq, based on the molar amount of the polymerization unit), 216A05 (57.3 mg, 0.12 mmol, 1.5 eq), PyBOP (64.0 mg, 0.12 mmol, 1.5 eq), DIPEA (0.09 mL, 0.49 mmol, 4.0 eq) and DMF (1.6 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 hours, and the complete conversion of the reactants was monitored by HPLC. When MTBE (10 mL) was added to DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was washed successively with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low-molecular-weight impurities were removed by ultrafiltration with a MeOH / aqueous solution. By HPLC monitoring, the low-molecular-weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain the product 216A17, 175 mg, white solid, purity 99.13%.

[0766] Example 60: Synthesis of Compound 216A18

[0767]

Chemical Structure

[0768] Synthesis of Compound 216A06: Under nitrogen protection at room temperature (25 °C), abiraterone (1.0 g, 2.86 mmol, 1.0 eq), acid anhydride (1.14 g, 8.6 mmol, 3.0 eq), DMAP (697 mg, 5.7 mmol, 2.0 eq), DIPEA (1.0 mL, 5.7 mmol, 2.0 eq) and anhydrous DCM (10 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h and monitored by TLC for the remaining starting materials. The solvent was spin-dried and separated by silica gel column chromatography (5 - 20% MeOH in DCM) to give the product 216A06, 0.95 g, white solid, in 69% yield.

[0769] 1 H NMR (400 MHz, CDCl3) δ 8.66 (s, 1H), 8.47 (d, J = 4.2 Hz, 1H), 7.74 (dt, J = 8.0, 1.9 Hz, 1H), 7.31 (dd, J = 8.0, 4.9 Hz, 1H), 6.09 ‐ 6.03 (m, 1H), 5.38 (d, J = 5.1 Hz, 1H), 4.73 ‐ 4.60 (m, 1H), 3.43 (s, 2H), 3.40 (s, 2H), 2.41 ‐ 2.20 (m, 3H), 2.11 ‐ 2.00 (m, 3H), 1.93 ‐ 1.42 (m, 9H), 1.15 ‐ 0.99 (m, 8H). Synthesis of Compound 216A18: Under nitrogen protection at room temperature (25 °C), 101T19 (200 mg, 0.082 mmol, 1.0 eq, based on the molar amount of the polymerization unit), 216A06 (59.2 mg, 0.12 mmol, 1.5 eq), PyBOP (64.0 mg, 0.12 mmol, 1.5 eq), DIPEA (0.09 mL, 0.49 mmol, 4.0 eq) and DMF (1.6 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reaction was monitored by HPLC. When MTBE (10 mL) was added to the DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was successively washed with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low-molecular-weight impurities were removed by ultrafiltration with an MeOH / aqueous solution. By HPLC monitoring, the low-molecular-weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain the product 216A18, 184 mg, white solid, purity 99.74%.

[0770] Example 61: Synthesis of Compound 216A26

[0771]

Chemical formula

[0772] Synthesis of Compound 216A24: Under nitrogen protection at room temperature (25 °C), abiraterone (1.05 g, 3.0 mmol, 1.0 eq), CDI (1.46 g, 9.0 mmol, 3.0 eq) and anhydrous DCM (15 mL) were added sequentially. The reaction mixture was stirred at room temperature for 2 h, and the complete conversion of the reaction was monitored by HPLC. When MTBE (50 mL) was added to the system, a white precipitate was formed. The solid was filtered, washed with MTBE, and dried with an oil pump to obtain the product 216A24, 1.15 g, white solid, yield 86%.

[0773] 11H NMR (400 MHz, CDCl3) δ 8.62 (d, J = 2.2 Hz, 1H), 8.46 (dd, J = 4.8, 1.6 Hz, 1H), 8.14 (s, 1H), 7.65 (dt, J = 7.9, 1.9 Hz, 1H), 7.43 (t, J = 1.5 Hz, 1H), 7.23 (dd, J = 7.9, 4.8 Hz, 1H), 7.07 (s, 1H), 6.04 ‐ 5.96 (m, 1H), 5.49 (d, J = 4.8 Hz, 1H), 4.92 ‐ 4.78 (m, 1H), 2.58 ‐ 2.49 (m, 2H), 2.33 ‐ 2.24 (m, 1H), 2.14 ‐ 2.03 (m, 4H), 1.99 ‐ 1.92 (m, 1H), 1.81 ‐ 1.46 (m, 8H), 1.23 ‐ 1.12 (m, 4H), 1.06 (s, 3H). Synthesis of Compound 216A20: Under nitrogen protection at room temperature (25 °C), Boc-10106 (0.96 g, 2.0 mmol, 1.0 eq), 216A24 (1.07 g, 2.4 mmol, 1.2 eq), KOH (22 mg, 0.4 mmol, 0.2 eq), and toluene (50 mL) were added sequentially. The reaction was stirred at 60 °C for 15 hours. HPLC was used to monitor the reaction where a large amount of starting materials still remained. The solvent was spin-dried and separated by silica gel column chromatography (5%-25% MeOH in DCM) to obtain 165 mg of the product 216A20 as a white solid, with a yield of 10%.

[0774] 11H NMR (400 MHz, CDCl3) δ 9.20 (s, 1H), 8.61 (d, J = 2.3 Hz, 1H), 8.45 (dd, J = 4.8, 1.7 Hz, 1H), 7.65 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 8.3 Hz, 2H), 7.32 (d, J = 8.4 Hz, 2H), 7.22 (dd, J = 8.0, 4.8 Hz, 1H), 7.14 (d, J = 8.0 Hz, 1H), 6.03 ‐ 5.95 (m, 1H), 5.43 (d, J = 5.1 Hz, 1H), 5.23 (t, J = 6.2 Hz, 1H), 5.18 (d, J = 8.1 Hz, 1H), 5.09 (s, 2H), 4.80 ‐ 4.70 (m, 1H), 4.63 (s, 2H), 4.56 ‐ 4.42 (m, 1H), 4.03 ‐ 3.93 (m, 1H), 3.49 ‐ 3.37 (m, 1H), 3.28 ‐ 3.15 (m, 1H), 2.45 ‐ 1.84 (m, 13H), 1.43 (s, 10H), 1.05 (d, J = 9.2 Hz, 6H), 0.98 (d, J = 6.7 Hz, 3H), 0.94 (d, J = 6.8 Hz, 3H). Synthesis of Compound 216A25: At room temperature (25 °C), TFA (0.2 mL) was added to a DCM solution (2 mL) of 216A20 (165 mg, 0.19 mmol, 1.0 eq). The reaction was stirred at room temperature for 0.5 h. When the reaction was monitored by HPLC, a small amount of starting material remained less than 10%, and a small amount of abiraterone due to the decomposition of the carbonate was detected. DCM (20 mL) and saturated sodium bicarbonate (20 mL) were added to the system, the organic phase was separated and collected, and the aqueous phase was washed three times with DCM solution. The combined organic phases were washed with saturated aqueous sodium chloride solution, the solvent was spin-dried, and separated by silica gel column chromatography (5% - 30% MeOH in DCM) to obtain the product 216A25, 84 mg, white solid, yield 59%.

[0775] 11H NMR (400 MHz, CD3OD) δ 8.55 (d, J = 2.3 Hz, 1H), 8.40 (dd, J = 4.9, 1.6 Hz, 1H), 7.89 ‐ 7.82 (m, 1H), 7.65 ‐ 7.56 (m, 2H), 7.42 ‐ 7.33 (m, 3H), 6.13 ‐ 6.07 (m, 1H), 5.48 (d, J = 5.1 Hz, 1H), 5.41 ‐ 5.32 (m, 1H), 5.11 (s, 2H), 4.61 ‐ 4.53 (m, 4H), 4.50 ‐ 4.38 (m, 1H), 3.27 ‐ 3.10 (m, 3H), 2.47 ‐ 2.28 (m, 3H), 2.22 ‐ 1.51 (m, 20H), 1.21 ‐ 1.08 (m, 8H), 1.01 (d, J = 6.9 Hz, 3H), 0.95 (d, J = 6.8 Hz, 3H). Synthesis of Compound 216A26: Under nitrogen protection at room temperature (25 °C), 101B13 (90 mg, 0.036 mmol, 1.0 eq, based on the molar amount of the polymerization unit), 216A25 (40.5 mg, 0.054 mmol, 1.5 eq), PyBOP (27.9 mg, 0.054 mmol, 1.5 eq), DIPEA (0.025 mL, 0.14 mmol, 4.0 eq) and DMF (0.8 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reactants was monitored by HPLC. When MTBE (10 mL) was added to DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was washed successively with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low molecular weight impurities were removed by ultrafiltration with MeOH / aqueous solution. By HPLC monitoring, the low molecular weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain the product 216A26, 120 mg, white solid, purity 97.13%.

[0776] Example 55: Synthesis of Compound 216B06

[0777]

Chemical Structure

[0778] Synthesis of Compound 216B01: While sequentially adding fulvestrant (930 mg, 1.53 mmol, 1.0 eq), DIPEA (0.41 mL, 2.3 mmol, 1.5 eq), and anhydrous DCM (15 mL), the reaction was carried out under nitrogen protection at -20 °C. The components were gently stirred to dissolve, and acyl chloride (433 mg, 2.15 mmol, 1.4 eq) was added. The reaction mixture was slowly returned to room temperature and stirred for 2 hours. The complete conversion of the reaction was monitored by HPLC. The reaction solution was used without treatment and directly proceeded to the next step.

[0779] Synthesis of Compound 216B02: Amine (584 mg, 3.1 mmol, 2.0 eq) was added to the above reaction solution and stirred at room temperature for 15 hours. The solvent was spin-dried and separated by silica gel column chromatography (0% - 10% MeOH in DCM) to obtain the product 216B02, 1.24 g, white solid, with a 99% yield in two steps.

[0780] 1 H NMR (400 MHz, CDCl3) δ 7.26 (d, J = 7.4 Hz, 1H), 6.91 ‐ 6.83 (m, 1H), 6.80 (s, 1H), 3.74 (t, J = 8.5 Hz, 1H), 3.63 ‐ 3.39 (m, 4H), 3.13 ‐ 2.99 (m, 3H), 2.95 ‐ 2.85 (m, 4H), 2.80 ‐ 2.69 (m, 4H), 2.69 ‐ 2.58 (m, 1H), 2.38 ‐ 2.09 (m, 7H), 1.97 ‐ 1.86 (m, 1H), 1.80 ‐ 1.58 (m, 9H), 1.52 ‐ 1.16 (m, 32H), 0.77 (s, 3H). Synthesis of Compound 216B03: TFA (1 mL) was added to a DCM solution (5 mL) of 216B02 (500 mg, 0.61 mmol, 1.0 eq) at 0 °C. The reaction mixture was stirred at room temperature for 1 hour, and the complete conversion of the reactants was monitored by HPLC. The solution was spin-dried to obtain a white solid.

[0781] Synthesis of compound 216B04: Under nitrogen protection at room temperature (25 °C), 10107 (702 mg, 0.92 mmol, 1.5 eq) and DIPEA (0.33 mL, 1.83 mmol, 3.0 eq) were added to a DMF solution (5 mL) of 216B03. DIPEA (0.2 mL) and 10107 (500 mg) were added, and the incomplete conversion of the reaction mixture stirred overnight was monitored by HPLC, and the complete conversion of the reactants was monitored by HPLC. The solvent was spin-dried and separated by silica gel column chromatography (0% - 15% MeOH in DCM) to obtain the product 216B04, 0.56 g, white solid, in a two-step process with a 68% yield over two steps.

[0782] Synthesis of compound 216B05: Under nitrogen protection at room temperature (25 °C), diethylamine (0.3 mL, 3.1 mmol, 5.0 eq) was added to a DMF solution of 216B04 (809 mg, 0.61 mmol, 1.0 eq). The reaction mixture was stirred at room temperature for 15 hours, and the complete conversion of the reactants was monitored by HPLC. The solvent was spin-dried and separated by silica gel column chromatography (5% - 30% MeOH in DCM) to obtain the product 216B05, 427 mg, white solid, with a 62% yield.

[0783] 11H NMR (400 MHz, CDCl3) δ 9.66 (s, 1H), 8.28 (s, 1H), 7.75 ‐ 7.45 (m, 2H), 7.25 ‐ 7.03 (m, 2H), 6.76 (d, J = 10.3 Hz, 2H), 5.88 (s, 1H), 5.29 ‐ 4.92 (m, 4H), 4.77 (s, 1H), 3.71 (s, 1H), 3.65 ‐ 2.56 (m, 24H), 2.58 ‐ 1.99 (m, 22H), 1.95 ‐ 1.09 (m, 38H), 1.04 ‐ 0.71 (m, 13H). Synthesis of Compound 216B06: Under nitrogen protection at room temperature (25 °C), 101B13 (150 mg, 0.06 mmol, 1.0 eq, based on the molar amount of the polymerization unit), 216B05 (101.4 mg, 0.09 mmol, 1.5 eq), PyBOP (46.8 mg, 0.09 mmol, 1.5 eq), DIPEA (0.04 mL, 0.24 mmol, 4.0 eq) and DMF (0.6 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reactants was monitored by HPLC. When MTBE (10 mL) was added to DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was successively washed with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low-molecular-weight impurities were removed by ultrafiltration with a MeOH / aqueous solution. By HPLC monitoring, the low-molecular-weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain the product 216B06, 200 mg, white solid, purity 98.76%.

[0784] Example 56: Synthesis of Compound 216D06:

[0785] [Chemical formula]

[0786] Synthesis of Compound 216D01 Under nitrogen protection at room temperature (25 °C), 101B13 (500 mg, 0.21 mmol, 1.0 eq, based on the molar amount of the polymerization unit), amine hydrochloride (62.6 mg, 0.32 mmol, 1.5 eq), PyBOP (165.0 mg, 0.32 mmol, 1.5 eq), DIPEA (0.19 mL, 1.06 mmol, 5.0 eq) and DMF (4.0 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reactants was monitored by HPLC. When MTBE (40 mL) was added to the DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was washed successively with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low molecular weight impurities were removed by ultrafiltration with a MeOH / aqueous solution. By HPLC monitoring, the low molecular weight impurities were completely removed. The MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain the product 216D01, 455 mg, as a white solid.

[0787] Synthesis of Compound 216D03 At room temperature (25 °C), 216D01 (277 mg, 0.11 mmol, 1.0 eq) and NaOH solution (0.1 M, 5.6 ml, 0.56 mmol, 5.0 eq) were added sequentially. The reaction mixture was stirred at room temperature for 4 h, and the complete conversion of the reactants was monitored by HPLC. This solution was ultrafiltered with water to remove low molecular weight impurities and lyophilized to obtain the product 216D03, 244 mg, as a white solid.

[0788] Synthesis of Compound 216D04 At room temperature (25 °C), an aqueous solution (10 ml) of 1,2-cyclohexanediamine (550 mg, 4.82 mmol, 1.0 eq) was slowly added dropwise to an aqueous solution (12 ml) of K2PtCl4 (2.0 g, 4.82 mmol, 1.0 eq). The reaction was stirred at room temperature for 15 h. The solution was filtered to obtain an earthy yellow solid. This solid was washed successively with water and methanol and evacuated with an oil pump to obtain the product 216D04, 1.6 g, as an earthy yellow solid.

[0789] Synthesis of Compound 216D08 At room temperature of 25 °C, 216D04 (226.9 mg, 0.6 mmol, 1.0 eq), silver sulfate (187.1 mg, 0.6 mmol, 1.0 eq) and water (3.0 ml) were sequentially added to a brown vial. The reaction was stirred at room temperature for 24 hours. The solution was filtered to obtain a clear solution of 216D08 (0.2 M).

[0790] Synthesis of Compound 216D06 At room temperature of 25 °C, Ba(OH)2 solution (0.1 M, 0.22 ml, 0.022 mmol, 1.0 eq) was added to an aqueous solution (0.5 ml) of 216D03 (55 mg, 0.022 mmol, 1.0 eq). The reactants were stirred at room temperature for 10 minutes. To this solution, 216D08 solution (0.2 M, 0.5 ml, 0.11 mmol, 5.0 eq) was added. The reaction was stirred at room temperature for 15 hours. The complete conversion of the reactants was monitored by HPLC. It was filtered to remove insoluble matters, ultrafiltered with water to remove low molecular weight impurities, and freeze-dried to obtain a clear solution, giving 51 mg of the product 216D06, a white solid with a purity of 98.5%.

[0791] Example 57: Synthesis of Compound 216D13

[0792]

Chemical formula

[0793] Synthesis of Compound 216D02 Under nitrogen protection at room temperature (25 °C), 101B13 (500 mg, 0.21 mmol, 1.0 eq), amine hydrochloride (67.1 mg, 0.32 mmol, 1.5 eq), PyBOP (165.0 mg, 0.32 mmol, 1.5 eq), DIPEA (0.19 mL, 1.06 mmol, 5.0 eq) and DMF (4.0 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 h, and the complete conversion of the reactants was monitored by HPLC. When MTBE (40 mL) was added to the DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was successively washed with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low molecular weight impurities were removed by ultrafiltration with a MeOH / aqueous solution. By HPLC monitoring, the low molecular weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain the product 216D02, 529 mg, as a white solid.

[0794] Synthesis of Compound 216D07 At room temperature (25 °C), 216D02 (250 mg, 0.1 mmol, 1.0 eq) and NaOH solution (0.1 M, 5.0 ml, 0.5 mmol, 5.0 eq) were added sequentially. The reaction mixture was stirred at room temperature for 3 h, and the complete conversion of the reactants was monitored by HPLC. This solution was ultrafiltered with water to remove low molecular weight impurities and lyophilized to obtain the product 216D07, 209 mg, as a white solid.

[0795] Synthesis of Compound 216D13 An aqueous solution (0.5 ml) of 216D07 (75 mg, 0.03 mmol, 1.0 eq) at room temperature (25 °C) was added to a Ba(OH)2 solution (0.1 M, 0.3 ml, 0.03 mmol, 1.0 eq). The reaction mixture was stirred at room temperature for 10 min. To this solution was added a 216D08 solution (0.2 M, 0.75 ml, 0.15 mmol, 5.0 eq). The reaction was stirred at room temperature for 15 h. The complete conversion of the reactants was monitored by HPLC. The insoluble matter was removed by filtration, and low molecular weight impurities were removed by ultrafiltration with water, followed by lyophilization to obtain a clear solution, and the product 216D13, 57 mg, as a white solid with a purity of 99%.

[0796] Example 58: Synthesis of Compound 216D36

[0797]

Chemical Structure

[0798] Synthesis of Compound 216D34 At room temperature (25 °C) under nitrogen protection, 101B13 (500 mg, 0.21 mmol, 1.0 eq, based on the molar amount of the polymerization unit), amine (176.8 mg, 0.32 mmol, 1.5 eq), PyBOP (165.0 mg, 0.32 mmol, 1.5 eq), DIPEA (0.19 mL, 1.06 mmol, 5.0 eq) and DMF (4.0 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 hours, and the complete conversion of the reaction was monitored by HPLC. When MTBE (40 mL) was added to the DMF, the solution became turbid with the precipitation of a white solid. The upper liquid layer was aspirated, and the white solid was washed successively with MTBE and aspirated. The remaining white solid was dissolved in MeOH, and low molecular weight impurities were removed by ultrafiltration with a MeOH / aqueous solution. By HPLC monitoring, the low molecular weight impurities were completely removed. MeOH was spin-off to recover the solution, and the aqueous solution was lyophilized to obtain 514 mg of the product 216D34 as a white solid.

[0799] Synthesis of Compound 216D35 At room temperature (25 °C), 216D34 (290 mg, 0.1 mmol, 1.0 eq) and NaOH solution (0.1 M, 5.0 ml, 0.5 mmol, 5.0 eq) were added sequentially. The reaction mixture was stirred at room temperature for 3 hours, and the complete conversion of the reaction was monitored by HPLC. This solution was ultrafiltered with water to remove low molecular weight impurities and lyophilized to obtain 245 mg of the product 216D35 as a white solid.

[0800] Synthesis of Compound 216D36 An aqueous solution (0.5 ml) of 216D35 (87 mg, 0.03 mmol, 1.0 eq) at room temperature (25 °C) was added to a Ba(OH)₂ solution (0.1 M, 0.3 ml, 0.03 mmol, 1.0 eq). The reaction mixture was stirred at room temperature for 10 minutes. A 216D08 solution (0.2 M, 0.75 ml, 0.15 mmol, 5.0 eq) was added to this solution. The reaction was stirred at room temperature for 15 hours. The complete conversion of the reactants was monitored by HPLC. The mixture was filtered to remove insoluble substances, ultrafiltered with water to remove low molecular weight impurities, and lyophilized to obtain a clear solution, yielding 67 mg of the product 216D36 as a white solid with a purity of 98%.

[0801] Example 59: Synthesis of Compound 21401

[0802]

Chemical Structure

[0803] Synthesis of Compound 21401F To a DCM solution (40 mL) of tert-butyl acetate (667 mg, 5.05 mmol, 2.0 eq), EDCI (965 mg, 5.05 mmol, 2.0 eq), and DMAP (615 mg, 5.05 mmol, 2.0 eq) was added 21401B (2.5 g, 2.52 mmol, 1.0 eq) at 0 °C. The reaction solution was stirred and reacted for 16 hours. As a result of LCMS, a highly converted product was formed. After confirming the completion of the reaction, the reaction solution was spin-dried and passed through a column with PE / EA = 9:1 to obtain 1.64 g of a white solid. MS (ESI), m / z, 1126.9 [M+Na] + ; Synthesis of Compound 21401G TFA (10 mL) was added dropwise to a DCM solution (10 mL) of 21401F (1.52 g, 1.38 mmol, 1.0 eq) at 0 °C. After the addition, the mixture was returned to room temperature and stirred for 2 hours. After the reaction was completed, the reaction solution was spin-dried. After the reaction was completed, the reaction solution was spin-dried, acetonitrile (200 mL) was added, and spin-distilled to remove residual TFA, yielding 1.6 g of a yellow solid.

[0804] MS (ESI), m / z, 949.0 [M+H] + ; m / z, 990.0 [M+H+CH3CN] + ; Synthesis of Compound 21401H At room temperature (25 °C), Boc2O (361 mg, 1.65 mmol, 1.2 eq) and Et3N (557 mg, 5.52 mmol, 4.0 eq) were sequentially added to a DCM solution (15 mL) of 21401G (1.52 g, 1.38 mmol, 1.0 eq). The reaction was stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was spin-dried and purified by a C18 column using CH3CN:H2O = 5:95 as the eluent. However, when purified by the C18 column, a part of the crude product deteriorated (in the previous batch, it could be purified without deterioration using a reverse-phase column. This is probably because a very small amount of residue of the packing material of the reverse-phase column induced the deterioration of the product), and only 230 mg of the product was obtained. MS (ESI), m / z, 1070.9 [M+Na] + ; Synthesis of Compound 21401J At room temperature (25 °C), TBAF (1 M in THF, 2.0 mL, 2.0 mmol, 2.9 eq) and AcOH (228 mg, 3.79 mmol, 5.5 eq) were sequentially added to a THF solution (20 mL) of 21401H (720 mg, 0.686 mmol, 1.0 eq). The reaction was stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was spin-dried. The crude product was first passed through a silica gel column, (PE / EA = 2:1~0:1)~DCM / MeOH = 10:1 to recover the product. The obtained crude product was further purified by a C18 column using CH3CN:H2O = 5~70% as the eluent, and 230 mg of the product was obtained. MS (ESI), m / z, 957.0 [M+Na]+.

[0805] Synthesis of Compound 21401I At room temperature of 25 °C, 21401H (50 mg, 0.048 mmol, 1.5 eq), PyBop (25 mg, 0.048 mmol, 1.50 eq) and DIPEA (17 mg, 0.128 mmol, 4.0 eq) were sequentially added to a DMF solution (2 mL) of 110T19 (78 mg, 0.032 mmol, 1.0 eq). After the addition, the mixture was stirred at room temperature overnight. The complete conversion of the reactants was monitored by HPLC. It was filtered to remove insoluble substances, ultrafiltered with water and methanol to remove low-molecular-weight impurities, and freeze-dried to obtain a clear solution, and the product 21401I (60 mg, white solid, purity 98.6%) was obtained.

[0806] Synthesis of Compound 21401 At room temperature of 25 °C, TFA (2 mL) was added dropwise to a CH2Cl2 solution (2 mL) of 21401I (60 mg). It was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was spin-dried. Saturated NaHCO3(aq) was added to adjust the pH to 7. The crude product was dissolved in H2O / MeOH (50 / 50 mL), ultrafiltered through a 30K MWCO ultrafiltration membrane, and freeze-dried to obtain 45 mg of a pale yellow solid.

[0807] Example 60: Synthesis of Compound 21404

[0808]

Chemical Structure

[0809] Synthesis of Compound 21401A To a suspension of 214M01 (5.0 g, 6.45 mmol, 1.0 eq) in dioxane / H2O solution (100 mL / 100 mL), Boc2O (1.55 g, 7.10 mmol, 1.1 eq) was added at 0 °C. TEA (980 mg, 9.70 mmol, 1.5 eq) was slowly added dropwise. After the dropwise addition, the reaction mixture was returned to room temperature and stirred for 16 h. After completion of the reaction, the reaction solution was poured into saturated NaCl (100 mL), partitioned, and the aqueous phase was extracted with EA (100 mL × 3). The combined organic phases were washed with saturated brine (500 mL×2), dried over anhydrous Na2SO4, and spun down to about 10 - 20 mL. The residue was added dropwise to acetonitrile (200 mL) and triturated at room temperature. The filter was drawn, and the cake was dried thoroughly to obtain 5.1 g of a light gray solid. The yield was 96.5%. MS (ESI), m / z, 821.10 [M+H-C4H8]; m / z, 777.10 [M+H-Boc] + ; m / z, 777.10 [M+H-Boc] + ; Synthesis of Compound 21401B TBDMSCl (3.44 g, 22.8 mmol, 8.0 eq) and TEA (1.15 g, 11.41 mmol, 4.0 eq) were continuously added dropwise to a DMF solution (20 mL) of 21401A (2.5 g, 2.85 mmol, 1.0 eq) at 0 °C. The reaction mixture was returned to room temperature and stirred for 2 h. After completion of the reaction, the reaction solution was poured into saturated NaHCO3(aq) (200 mL), quenched, and extracted with CH2Cl2 (150 mL × 2). The combined organic phases were washed with saturated brine, dried over anhydrous Na2SO4, and spin-dried. The crude product was passed through a reverse-phase column with CH3CN:H2O = 5 - 95% to obtain 2.5 g of a white solid. MS (ESI), m / z, 936.20 [M+H-C4H8]+.

[0810] Synthesis of Compound 21401C At room temperature (25 °C), Boc2O (593 mg, 2.72 mmol, 2.0 eq) and DMAP (183 mg, 1.50 mmol, 1.1 eq) were added to a solution of 21401B (1.5 g, 1.36 mmol, 1.0 eq) in tert-butanol (20 mL). The reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was spin-dried and passed through a silica gel column (PE / EA = 20:1), and 1.21 g of a white solid was obtained. MS (ESI), m / z, 991.10 [M+H-C4H8] + .

[0811] Synthesis of Compound 21401D At room temperature (25 °C), TBAF (1 M, 2.9 mL, 2.9 mmol, 2.5 eq) and AcOH (345 mg, 5.75 mmol, 5.0 eq) were added dropwise to a solution of 21404C (1.2 g, 1.15 mmol, 1.0 eq) in THF (25 mL), and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was spin-dried, and first passed through a silica gel column using PE / EA = 4:1 as the eluent to obtain the crude product. Next, the crude product was passed through a reverse phase of CH3CN:H2O = 5 - 70%, and 1.05 g of a pale yellow solid was obtained. MS (ESI), m / z, 876.30 [M+H-C4H8] + .

[0812] Synthesis of Compound 21404E At room temperature (25 °C), to a solution of 21401D (1.05 g, 1.13 mmol, 1.0 eq) in CH₂Cl₂, glutaric anhydride (386 mg, 3.39 mmol, 3.0 eq), DIPEA (583 mg, 4.52 mmol, 4.0 eq) and DMAP (551 mg, 4.52 mmol, 4.0 eq) were sequentially added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was poured into saturated aqueous citric acid (50 mL), partitioned, and the aqueous phase was extracted with CH₂Cl₂ (100 mL × 3). The combined organic phases were washed with saturated brine (500 mL × 2), dried over anhydrous Na₂SO₄, spin-dried, passed through a column with PE / EA = 1:1 to DCM / MeOH = 10:1, and 700 mg of a white solid was obtained. MS (ESI), m / z, 935.00 [M+H-C₄H₈-C₄H₈]+.

[0813] 1 ¹H NMR (400 MHz, Chloroform-d) δ 7.71 (s, 2H), 7.21 (s, 2H), 5.20 (d, J = 7.6 Hz, 1H), 4.44 (d, J = 6.9 Hz, 1H), 3.15 ‐ 2.91 (m, 2H), 2.80 (t, J = 7.2 Hz, 2H), 2.63 (t, J = 7.3 Hz, 2H), 2.19 (p, J = 7.3 Hz, 2H), 1.49 (s, 9H), 1.47 (s, 9H). Synthesis of Compound 21404F At room temperature (25 °C), to a solution of 110T19 (390 mg, 0.16 mmol, 1.0 eq) in DCM (5 mL), 21404E (250 mg, 0.238 mmol, 1.49 eq), PyBop (125 mg, 0.240 mmol, 1.50 eq) and DIPEA (85 mg, 0.657 mmol, 4.10 eq) were sequentially added. After addition, the mixture was stirred at room temperature overnight. MTBE (50 mL) was added and triturated, and stirred for 10 minutes. Left standing for a long time, the supernatant was poured off, and the residual solid was dried in vacuo to obtain 610 mg of the crude product.

[0814] Synthesis of Compound 21404 At room temperature of 25 °C, TFA (3 mL) was added dropwise to a CH₂Cl₂ solution (3 mL) of 21404E (610 mg, 0.185 mmol, 1.0 eq). The mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was spin-dried. Saturated NaHCO₃(aq) was added to adjust the pH to 7. The crude product was dissolved in H₂O / MeOH (50 / 50 mL), ultrafiltered through a 30K MWCO ultrafiltration membrane, and lyophilized to obtain 250 mg of a pale yellow solid.

[0815] Example 61: Synthesis of Compound 21701

[0816]

Chemical Structure

[0817] Synthesis of Compound 21701-1 Under nitrogen protection at room temperature of 25 °C, DIPEA (1.14 g, 8.82 mmol, 3.0 eq) was added to a DMF (60 mL) solution of octreotide acetate (3.0 g, 2.94 mmol, 1.0 eq). The reaction solution was cooled to -40 °C, and a solution of tert-butyl N-succinimidyl carbonate (633 mg, 2.94 mmol, 1.0 eq) in DMF (10 mL) was added dropwise slowly. The reaction was terminated by LCMS detection, and the reaction solution was purified by silica gel column to obtain product 5 as a white solid (2.8 mg, yield 85%). MS (ESI), m / z, 1118.40[M+H]+.

[0818] Synthesis of Compound 21701-2 Under room temperature and nitrogen protection, to the DMF (10 mL) of SM-1 (102 mg, 0.13 mmol, 1.0 eq), 21701-1 (150 mg, 0.13 mmol, 1.0 eq), HOBT (35 mg, 0.26 mmol, 2.0 eq) and 2,6-dimethylpyridine (43 mg, 0.40 mmol, 3.0 eq) were added. Then, it was stirred overnight. After the reaction was completed, it was diluted with 50 ml of water, filtered, dried, and 220 mg of a white solid was obtained with a yield of 96.9%. MS (ESI), m / z, 1746.2 [M+H] + 。

[0819] Synthesis of Compound 21701-3 Under room temperature and nitrogen protection, to the dichloromethane / methanol (5:1, 10 mL) of 21701-2 (270 mg, 0.15 mmol, 1.0 eq), piperidine (20 mg, 0.23 mmol, 1.5 eq) was added. Then, it was stirred overnight. After the reaction was completed, it was concentrated directly and purified by column chromatography separation to obtain 210 mg of a white solid with a yield of 89%. MS (ESI), m / z, 1524.2 [M+H]+. Synthesis of Compound 21701-4.

[0820] Under room temperature of 25°C and nitrogen protection, to the DMF (10 mL) of 21701-3 (0.1 g, 0.065 mmol, 1.8 eq), 101B13 (0.09 g, 0.036 mmol, based on the molar amount of the polymerization unit), DIEA (0.2 mL) and PyBOP (0.037 g, 0.072 mmol) were added. Then, it was stirred overnight. The reaction was monitored by HPLC, and when the reaction was completed, the crude product was concentrated and purified to 150 mg and used directly in the next step.

[0821] Synthesis of Compound 21701 At room temperature (0 °C) under nitrogen protection, HCl / Dioxane (4 M, 0.5 mL) was added to 21701-4 (0.15 g) in DCM / MeOH (5:1, 10 mL), and the reaction was monitored by HPLC. After completion of the reaction, 90 mg of the product was obtained by ultrafiltration (30 K MW, methanol and water), freeze-drying and purification, purity: 98.79%.

[0822] Example 62: Synthesis of Compound 21702

[0823]

Chemical Structure

[0824] Synthesis of Compound 21702-2 At room temperature under nitrogen protection, 21701-1 (300 mg, 0.29 mmol, 1.0 eq), HOBT (80 mg, 0.58 mmol, 2.0 eq) and 2,6-dimethylpyridine (95 mg, 0.87 mmol, 3.0 eq) were added to SM-1 (225 mg, 0.29 mmol, 1.0 eq) in DMF (10 mL). Then, it was stirred overnight. After completion of the reaction, it was diluted with 50 ml of water, filtered, dried, and 310 mg of a white solid was obtained in a yield of 63.9%. MS (ESI), m / z, 1646.2 [M+H] + .

[0825] Synthesis of Compound 21702-3 At room temperature under nitrogen protection, Boc2O (50 mg, 0.23 mmol, 1.2 eq) and a catalytic amount of DMAP were added to 21702-2 (310 mg, 0.188 mmol, 1.0 eq) in dichloromethane (20 mL). Then, it was stirred overnight. After completion of the reaction, it was concentrated as it was and purified by column chromatography separation, and 250 mg of a white solid was obtained in a yield of 76%. MS (ESI), m / z, 1746.2 [M+H] + .

[0826] Synthesis of Compound 21702-4 Under nitrogen protection at room temperature, piperidine (20 mg, 0.23 mmol, 1.5 eq) was added to dichloromethane / methanol (5:1, 10 mL) of 21701-3 (250 mg, 0.14 mmol, 1.0 eq). Then, it was stirred overnight. After completion of the reaction, it was concentrated as it was, and 130 mg of a white solid with a yield of 59.6% was obtained by pre-HPLC. MS (ESI), m / z, 1524.2 [M+H] + 。

[0827] Synthesis of Compound 21702-5 Under nitrogen protection at room temperature (25 °C), 101B13 (0.119 g, 0.047 mmol), DIEA (0.2 mL), and PyBOP (0.037 g, 0.072 mmol) were added to DMF (10 mL) of 21702-4 (0.13 g, 0.085 mmol, 1.8 eq). Then, it was stirred overnight. The reaction was monitored by HPLC, and when the reaction was completed, the crude product was concentrated and purified to 160 mg and used as it was in the next step.

[0828] Synthesis of Compound 21702 Under nitrogen protection at 0 °C at room temperature, HCl / Dioxane (4M, 0.5 mL) was added to DCM / MeOH (5:1, 10 mL) of 21702-5 (0.16 g,), and the reaction was monitored by HPLC. At the end of the reaction, the product was obtained by ultrafiltration (30KMW), freeze-drying, and purification, and 65 mg of the product was obtained, purity: 98%.

[0829] Example 70: Synthesis of Compound 21703

[0830]

Chemical Structure

[0831] Synthesis of Compound 21703-2 Under nitrogen protection at room temperature, 21701-1 (160 mg, 0.143 mmol, 1.0 eq), HOBT (40 mg, 0.29 mmol, 2.0 eq) and 2,6-dimethylpyridine (48 mg, 0.42 mmol, 3.0 eq) were added to DMF (10 mL) of SM-1 (50 mg, 0.143 mmol, 1.0 eq). Then, it was stirred overnight. After completion of the reaction, it was diluted with 50 ml of water, filtered and dried to obtain 130 mg of a white solid with a yield of 68%. MS (ESI), m / z, 1332.2 [M+H] + 。

[0832] Synthesis of Compound 21703-3 Under nitrogen protection at room temperature, SM-2 (20 mg, 0.108 mmol, 1.2 eq) and a catalytic amount of acetic acid were added to DMF (10 mL) of 21702-2 (120 mg, 0.09 mmol, 1.0 eq). Then, it was stirred overnight. After completion of the reaction, it was concentrated as it was and purified by preparative-HPLC to obtain 60 mg of a white solid with a yield of 46.5%. MS (ESI), m / z, 1369.2 [M+H] + 。

[0833] Synthesis of Compound 21703 Under nitrogen protection at 25 °C (room temperature), 101T19 (69 mg, 0.029 mmol, based on the molar amount of the polymerization unit), DIEA (0.2 mL) and PyBOP (0.037 g, 0.072 mmol) were added to DMF (10 mL) of 21703-3 (60 mg, 0.043 mmol, 1.5 eq). Then, it was stirred overnight. After completion of the reaction, the crude product was directly concentrated and purified to 80 mg, dissolved in DCM / MeOH (5:1, 10 mL), HCl / Dioxane (4M, 0.5 mL) was added, and the reaction was monitored by HPLC. After completion of the reaction, the product was ultrafiltered (30K MW), freeze-dried and purified to obtain 60 mg (purity: 99.5%).

[0834] Example 71: Synthesis of Compound 21704

[0835] [Chemistry]

[0836] Synthesis of Compound 21704-1 Under nitrogen protection at room temperature, SM-2 (133 mg, 1.15 mmol, 1.2 eq) and pyridine (230 mg, 2.88 mmol, 3.0 eq) were added to dichloromethane (20 mL) of SM-1 (500 mg, 0.96 mmol, 1.0 eq). Then, it was stirred overnight. After the reaction was completed, it was concentrated directly and purified by preparative HPLC to obtain 400 mg of a white solid with a yield of 65.4%. MS (ESI), m / z, 638.2 [M+H] + 。

[0837] Synthesis of Compound 21704 Under nitrogen protection at 25 °C (room temperature), 101T19 (0.992 g, 0.418 mmol, based on the molar amount of the polymerization unit), DIEA (0.5 mL) and PyBOP (0.435 g, 0.836 mmol) were added to DMF (100 mL) of 21704-1 (0.4 g, 0.627 mmol, 1.5 eq). Then, it was stirred overnight. After the reaction was completed, ultrafiltration (30K MW), freeze-drying, and purification were carried out to obtain 920 mg of a product with a purity of 98%.

[0838] Example 63: Synthesis of Compound 21705

[0839] [Chemistry]

[0840] Synthesis of Compound 21705-1 Under room temperature and nitrogen protection, leuprorelin (750 mg, 0.62 mmol, 1.0 eq) and pyridine (147 mg, 1.86 mmol, 3.0 eq) were added to DMF (10 mL) of SM-1 (94 mg, 0.807 mmol, 1.3 eq). Then, it was stirred overnight. After the reaction was completed, it was diluted with 50 ml of water, filtered, and dried to obtain 260 mg of a white solid with a yield of 31.7%. MS (ESI), m / z, 1325.2 [M+H] + 。

[0841] Synthesis of Compound 21705 Under room temperature (25 °C) and nitrogen protection, 101T19 (119 mg, 0.05 mmol, based on the molar amount of the polymerization unit), DIEA (0.2 mL), and PyBOP (0.052 g, 0.1 mmol) were added to DMF (10 mL) of 21705-1 (100 mg, 0.075 mmol, 1.5 eq). Then, it was stirred overnight. At the end of the reaction, 95 mg of the product was obtained by ultrafiltration (30K MW), freeze-drying, and purification, and the purity was 99%.

[0842] Example 64: Synthesis of Compound 21706

[0843]

Chemical Structure

[0844] Synthesis of Compound 21706-1 Under room temperature and nitrogen protection, 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione (1.2 g, 6.1 mmol, 1.2 eq) and DIPEA (1.3 h, 10.1 mmol, 2.0 eq) were added to DMF (20 mL) of SM-1 (1.4 g, 5.1 mol, 1.0 eq). The mixture was heated to 90 °C and stirred for 3 hours. It was quenched with water, extracted with dichloromethane (100 mL × 4), the organic phases were combined, washed with salt, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 800 mg of product 21706-1, a pale yellow solid. LCMS (ESI) M-100 = 361.1; M-56 = 405.0 Synthesis of Compound 21706-2 Under an ice-water bath and nitrogen protection, a dioxane hydrochloride solution (4.0 M, 10 mL) was added to 21706-1 (0.8 g, 1.74 mmol) in dichloromethane (10 mL)), and it was stirred for 1 hour. The reaction solution was concentrated as it was and used in the next step. MS (ESI), m / z, 361.1 [M+1]+.

[0845] Synthesis of Compound 21706-3 Under room temperature and nitrogen protection, methyl bromoacetate (552 mg, 3.6 mmol, 1.2 eq) and potassium carbonate (500 mg, 3.6 mmol, 1.2 eq) (4.0 M, 10 mL) were added to DMF (10 mL) of 21706-3M (prepared according to Example 40 of Patent CN108794452B, 1.6 g, 3.0 mmol). It was heated to 50 °C and stirred for 1 hour. The reaction product was quenched with water, extracted with dichloromethane (100 mL × 4), the organic phases were combined, washed with salt, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 1.4 g of product 21706-3, a pale yellow solid. MS (ESI), m / z, 566.2 [M+1]+.

[0846] Synthesis of Compound 21706-4 Under an ice bath and nitrogen protection, lithium hydroxide (600 mg, 25 mmol) was added to a mixed solution of 21706-3 (1.4 g, 2.48 mmol) in THF (20 mL) and water (5), and the mixture was stirred for 2 hours. The reaction was quenched with water, extracted with dichloromethane (100 mL × 4), the organic phases were combined, washed with salt, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 1.3 g of the product 21706-4 as a pale yellow solid. MS (ESI), m / z, 552.1 [M+1]+.

[0847] Synthesis of Compound 21706-5 Under an ice bath and nitrogen protection, HATU (1.3 g, 3.3 mmol, 2.0 eq) and DIPEA (430 mg, 3.3 mmol, 2.0 eq) were added to a solution of 21706-2 (600 mg, 1.7 mmol, 1.0 eq) and 21706-4 (920 mg, 1.7 mmol, 1.0 eq) in DCM (20 mL), and the mixture was stirred for 2 hours. The reaction was quenched with water, extracted with dichloromethane (100 mL x 4), the organic phases were combined, washed with salt, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 1.1 g of the product 21706-5. MS (ESI), m / z, 894.2 [M+1]+.

[0848] 11H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 11.00 (s, 1H), 9.52 (s, 1H), 8.80 (d, J = 3.6 Hz, 1H), 8.62 (t, J = 5.5 Hz, 1H), 8.35 ‐ 8.24 (m, 2H), 8.03 (d, J = 8.7 Hz, 1H), 7.92 (dd, J = 8.9, 2.2 Hz, 1H), 7.73 (d, J = 11.9 Hz, 1H), 7.58 (dd, J = 8.6, 7.1 Hz, 1H), 7.13 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.58 (d, J = 5.8 Hz, 1H), 5.05 (dd, J = 12.7, 5.4 Hz, 1H), 4.86 (p, J = 6.9 Hz, 1H), 4.45 (d, J = 6.1 Hz, 1H), 3.85 ‐ 3.80 (m, 2H), 3.61 (t, J = 5.4 Hz, 2H), 3.47 (dq, J = 13.4, 8.6, 7.1 Hz, 6H), 3.32 (t, J = 5.8 Hz, 2H), 3.02 ‐ 2.82 (m, 3H), 2.67 (s, 3H), 2.63 ‐ 2.56 (m, 1H), 2.53 (d, J = 4.4 Hz, 1H), 2.46 (d, J = 4.3 Hz, 1H), 2.06 ‐ 1.98 (m, 1H), 1.90 (d, J = 13.2 Hz, 1H), 1.78 (s, 1H), 1.63 (d, J = 6.9 Hz, 6H), 1.60 ‐ 1.50 (m, 2H). Synthesis of Compound 21706-6 Under room temperature and nitrogen protection, diethylene glycol anhydride (78 mg, 0.68 mmol, 1.2 eq) and pyridine (132 mg, 1.68 mmol, 3.0 eq) were added to dichloromethane (10 mL) of 21706-5 (500 mg, 0.56 mmol, 1.0 eq), and the mixture was stirred for 12 hours. After the reaction was completed, it was concentrated directly and purified by Pre-HPLC separation to obtain 400 mg of a white solid. MS (ESI), m / z, 1010.3 [M+H] + 。

[0849] Synthesis of Compound 21706 Under room temperature (25 °C) and nitrogen protection, 21706-6 (0.38 g, 0.38 mmol, 1.5 eq), PyBOP (262 mg, 0.5 mmol, 2.0 eq) and DIPEA (147 mg, 1.1 mmol, 4.5 eq) were added to a DMF (10 mL) solution of Compound 101T19 (600 mg, 0.25 mmol, 1.0 eq, based on the molar amount of the polymerization unit). The mixture was stirred overnight. After the reaction was completed, a 700 mg product was obtained by ultrafiltration (30 K MWCO), freeze-drying and purification.

[0850] Example 65: Synthesis of Compound 20323C

[0851]

Chemical Structure

[0852] Synthesis of Compound 20323C-1 Under room temperature and nitrogen protection, Fmoc-Val-Cit-PAB-PNP (CAS No. 863971-53-3, 0.79 g, 1.04 mmol, 1.1 eq) and DIPEA (0.36 g, 2.82 mmol, 3.0 eq) were added to a DMF (10 mL) suspension of exatecan mesylate hydrate (0.5 g, 0.94 mmol, 1.0 eq). The mixture was stirred overnight, concentrated directly, and purified by column chromatography separation to obtain 0.85 g of 20323C-1 with a yield of 85%.

[0853] Synthesis of Compound 20323C-2 Under an ice-water bath and nitrogen protection, piperidine (1 mL) was added to a suspension of 20323C-1 (0.8 g, 0.75 mmol, 1.0 eq) in DMF (5 mL). The mixture was stirred for 1 hour, concentrated directly, and purified by column chromatography separation to obtain 20323C-2, 0.55 g, with a yield of 87%.

[0854] Synthesis of Compound 20323C Under nitrogen protection at room temperature (25 °C), 101B13 (800 mg, 0.01 mmol, 1.0 eq), 20323C-2 (420 mg, 0.50 mmol, 46.0 eq), PyBOP (351 mg, 0.68 mmol, 62.0 eq), DIPEA (108 mg, 0.84 mmol, 77.0 eq) and DMF (5 mL) were added sequentially. The reaction mixture was stirred at room temperature for 15 hours, and the complete conversion of the reactants was monitored by HPLC. MTBE (20 mL) was added to DMF and stirred for 30 minutes. The upper layer was removed by suction, the residue was dissolved in MeOH and water (1 / 1), and ultrafiltration was carried out with an MeOH / aqueous solution to remove low-molecular-weight impurities. The low-molecular-weight impurities were completely removed by HPLC monitoring, the solution was recovered, MeOH was spin-off, and the aqueous solution was lyophilized to obtain Compound 20323C, 880 mg, a white solid, with a purity of 98.1%.

[0855] Example 66: Synthesis of Compound 22101

[0856]

Chemical Structure

[0857] Synthesis of Compound 22101A Under an ice-water bath and nitrogen protection, to a solution of 22101SM1 (synthesis example of compound 7 in J. Med. Chem. 2000, 43, 3093 - 3102., 0.5 g, 0.49 mmol, 1.0 eq) in DMF (5 mL) were added 22101SM2 (TFA salt, synthesis example of Example 96 in Patent WO2016151432A1, 0.41 g, 0.49 mmol, 1.0 eq) and DIPEA (0.32 g, 2.46 mmol, 5.0 eq). The mixture was returned to room temperature, stirred overnight, concentrated as it was, and used for the next reaction. MS (ESI), m / z, 1595.2 [M + H] + 。

[0858] Synthesis of Compound 22101B Under an ice-water bath and nitrogen protection, piperidine (1 mL) was added to a solution of the above crude product 22101A in DMF (5 mL), and the mixture was stirred for 1 hour. It was concentrated as it was and purified by Prep-HPLC to obtain 450 mg of Compound 22101B. MS (ESI), m / z, 1373.6 [M + H] + 。

[0859] Synthesis of Compound 22101 At room temperature (25 °C) and under nitrogen protection, 101B13 (500 mg, 0.007 mmol, 1.0 eq), 22101B (430 mg, 0.31 mmol, 46.0 eq), PyBOP (220 mg, 0.42 mmol, 62.0 eq), DIPEA (68 mg, 0.52 mmol, 77.0 eq) and DMF (5 mL) were added sequentially. The reaction mixture was stirred at room temperature overnight, and the complete conversion of the reactants was monitored by HPLC. MTBE (20 mL) was added to DMF, and the mixture was stirred for 30 minutes. The upper layer was removed by suction, the residue was dissolved in MeOH and water (1 / 1), and ultrafiltration was performed with a MeOH / aqueous solution to remove low-molecular-weight impurities. The low-molecular-weight impurities were completely removed by HPLC monitoring, the solution was recovered, MeOH was spin-off, and the aqueous solution was lyophilized to obtain 750 mg of Compound 22101, a white solid with a purity of 98.8%.

[0860] Example 67: Synthesis of Compound 22102

[0861]

Chem.

[0862] Synthesis of Compound 22102A Under an ice-water bath and nitrogen protection, 22101SM2 (0.37 g, 0.44 mmol, 1.0 eq) and DIPEA (0.28 g, 2.20 mmol, 5.0 eq) were added to a DMF (5 mL) solution of 22102SM (Synthesis example of Compound 14 in the literature ChemPlusChem 2013, 78, 222 - 226., 0.5 g, 0.44 mmol, 1.0 eq). The mixture was returned to room temperature, stirred overnight, concentrated as it was, and used in the next reaction. MS (ESI), m / z, 1709.2 [M+H] + .

[0863] Synthesis of Compound 22102B Under an ice-water bath and nitrogen protection, piperidine (1 mL) was added to a DMF (5 mL) solution of the above crude product 22102A, and the mixture was stirred for 1 hour, concentrated as it was, and purified by Prep-HPLC to obtain 485 mg of Compound 22102B. MS (ESI), m / z, 1487.6 [M+H] + .

[0864] Synthesis of Compound 22102 Under nitrogen protection at room temperature (25 °C), to a DMF solution (5 mL) of 101B13 (500 mg, 0.007 mmol, 1.0 eq), 22102B (467 mg, 0.31 mmol, 46.0 eq), PyBOP (220 mg, 0.42 mmol, 62.0 eq), DIPEA (68 mg, 0.52 mmol, 77.0 eq) and DMF (5 mL) were sequentially added. The reaction mixture was stirred at room temperature overnight, and the complete conversion of the reactants was monitored by HPLC. MTBE (20 mL) was added to the DMF, and the mixture was stirred for 30 minutes. The upper liquid layer was aspirated, and the residue was solvated with 5 mL of dry dichloromethane, cooled in an ice-water bath, 1 mL of trifluoroacetic acid was added, and the mixture was stirred for 1 hour. The completeness of the reaction was monitored by HPLC, and then concentrated directly. The residue was purified by ultrafiltration (30K MWCO) to obtain 720 mg of compound 22102 as a white solid with a purity of 99.1%.

[0865] Example 68: Synthesis of Compound 22103

[0866]

Chemical Structure

[0867] Synthesis of Compound 22103A Under nitrogen protection in an ice-water bath, to a DMF (5 mol) solution of 22103SM (Synthesis Example of Compound 72 in Patent WO2015038649A1, 0.5 g, 0.51 mmol, 1.0 eq), 22101SM2 (0.43 g, 0.51 mmol, 1.0 eq) and DIPEA (0.33 g, 2.57 mmol, 5.0 eq) were added. The mixture was returned to room temperature and stirred overnight. The crude product was concentrated directly and used in the next reaction. MS (ESI), m / z, 1549.2 [M+H] + .

[0868] Synthesis of Compound 22103B Under an ice bath and nitrogen protection, piperidine (1 mL) was added to a DMF (5 mL) solution of the above crude product 22103A, and the mixture was stirred for 1 hour. It was concentrated as it was and purified by Prep-HPLC to obtain 550 mg of compound 22103B. MS (ESI), m / z, 1327.7 [M+H] + 。

[0869] Synthesis of Compound 22103 Under nitrogen protection at room temperature (25 °C), 101B13 (500 mg, 0.0068 mmol, 1.0 eq), 22103B (416 mg, 0.31 mmol, 46.0 eq), PyBOP (220 mg, 0.42 mmol, 62.0 eq), DIPEA (68 mg, 0.52 mmol, 77.0 eq) and DMF (5 mL) were added sequentially. The reaction mixture was stirred at room temperature overnight, and the complete conversion of the reactants was monitored by HPLC. MTBE (20 mL) was added to DMF and stirred for 30 minutes. The upper liquid layer was aspirated, and the residue was purified by ultrafiltration (30 K MWCO) to obtain 650 mg of compound 22103, a white solid with a purity of 97.9%.

[0870] Example 69: Synthesis of Compound 22104

[0871]

Chemical formula

[0872] Synthesis of Compound 22104A Under an ice bath and nitrogen protection, 22101SM2 (0.42 g, 0.50 mmol, 1.0 eq) and DIPEA (0.32 g, 2.50 mmol, 5.0 eq) were added to a DMF (5 mL) solution of 22104SM (Synthesis example of compound 20' in Patent WO2017180834A1, 0.5 g, 0.50 mmol, 1.0 eq). The mixture was returned to room temperature and stirred overnight, and the crude product was concentrated as it was and used in the next reaction.

[0873] MS (ESI), m / z, 1577.2 [M+H]+.

[0874] Synthesis of Compound 22104B Under an ice-water bath and nitrogen protection, piperidine (1 mL) was added to a DMF (5 mL) solution of the above crude product 22104A, and the mixture was stirred for 1 hour. It was concentrated as it was and purified by Prep-HPLC to obtain 570 mg of Compound 22104B. MS (ESI), m / z, 1355.7 [M+H] + 。

[0875] Synthesis of Compound 22104 At room temperature (25 °C) and under nitrogen protection, 101B13 (500 mg, 0.0068 mmol, 1.0 eq), 22104B (424 mg, 0.31 mmol, 46.0 eq), PyBOP (220 mg, 0.42 mmol, 62.0 eq), DIPEA (68 mg, 0.52 mmol, 77.0 eq) and DMF (5 mL) were added sequentially. The reaction mixture was stirred at room temperature overnight, and the complete conversion of the reactants was monitored by HPLC. MTBE (20 mL) was added to DMF, and the mixture was stirred for 30 minutes. The upper liquid layer was aspirated, and the residue was purified by ultrafiltration (30 K MWCO) to obtain 580 mg of Compound 22104, a white solid with a purity of 99.0%.

[0876] Example 70: Synthesis of Compound 22105

[0877]

Chemical Structure

[0878] Synthesis of Compound 22105A Under - 40 °C and nitrogen protection, to a dry dichloromethane (10 mL) solution of 22105SM (synthesis example of compound 3a in the literature Molecules 2011, 16, 6769 - 6777, 0.5 g, 0.48 mmol, 1.0 eq), pyridine (0.5 mL) and p - nitrophenyl chloroformate (0.48 g, 2.41 mmol, 5.0 eq) were added. The reaction solution was stirred for 1 hour and directly concentrated, then a DMF solution (5 mL) was added to dissolve it. Subsequently, 22101SM2 (0.48 g, 0.58 mmol, 1.2 eq) and DIPEA (0.31 g, 2.41 mmol, 5.0 eq) were added. The mixture was returned to room temperature and stirred overnight. The crude product was directly concentrated and purified by column chromatography separation to obtain 450 mg of compound 22105A. MS (ESI), m / z, 1781.1 [M + H] + 。

[0879] Synthesis of Compound 22105B Under the protection of an ice - water bath and nitrogen, piperidine (1 mL) was added to a DMF (5 mL) solution of compound 22105A (450 mg), and the mixture was stirred for 1 hour. It was concentrated and purified by Prep - HPLC separation to obtain 350 mg of compound 22105B. MS (ESI), m / z, 1559.7 [M + H] + 。

[0880] Synthesis of Compound 22105 Under nitrogen protection at room temperature (25 °C), 101B13 (300 mg, 0.0041 mmol, 1.0 eq), 22105B (293 mg, 0.19 mmol, 46.0 eq), PyBOP (132 mg, 0.25 mmol, 62.0 eq), DIPEA (41 mg, 0.32 mmol, 77.0 eq) and DMF (5 mL) were added sequentially. The reaction mixture was stirred at room temperature overnight, and the complete conversion of the reactants was monitored by HPLC. MTBE (20 mL) was added to the DMF and stirred for 30 minutes. The upper layer liquid was aspirated, and the residue was dissolved in THF (10 mL). An ice-water bath was used, TBAF (1.0 M, 1 mL) was added, and the pH was adjusted to 7 with acetic acid. The reaction was stirred for 12 hours. The reaction was monitored for completeness by HPLC, and the compound was purified directly by ultrafiltration (30K MWCO) to obtain 380 mg of compound 22105 as a white solid with a purity of 98.2%.

[0881] Biological Safety Evaluation Example A: Comparison of the DOX Enzyme Drug Release Rates of 101A1601 and 101B00A

[0882] [Table 11]

[0883] [Table 12]

[0884] The polymer solution with papain buffer was placed in a 37 °C water bath and shaken. The released polymer solution was taken out of the test tube at different time points, returned to room temperature, 0.500 mL was pipetted, and diluted with 0.500 mL of DMSO solution. The concentration of the released adriamycin was measured by HPLC, and the ratio of the released adriamycin was calculated.

[0885] [Table 13]

[0886] Over a 4-hour period, the average drug release rate of the ε-poly-L-lysine derivative-drug coupling 101B00A was 11.6 times higher than that of the dendrimer 101A1601 (PEG1K) (Figure 1), indicating that the ε-poly-L-lysine derivative-drug coupling with lower site resistance enables a faster drug release rate.

[0887] Biological Safety Evaluation Example B: Comparison of the DOX Enzyme Drug Release Rates of 101A1602 and 101B00B

[0888] [Table 14]

[0889] [Table 15]

[0890] The polymer solution added with papain buffer was placed in a 37-degree water bath and shaken. The released polymer solution was taken out from the test tube at different time points, returned to room temperature, 0.500 mL was pipetted, and diluted with 0.500 mL of DMSO solution. The concentration of the released adriamycin was measured by HPLC, and the ratio of the released adriamycin was calculated.

[0891] [Table 16]

[0892] The average drug release rate of the ε-poly-L-lysine derivative-drug coupling 101B00B over 4 hours was 1.5 times that of the dendrimer 101A1602 (PEG 2K ), which indicates that PEG 2kEven when coupled, ε-poly-L-lysine derivative-drug coupling has shown to have lower steric resistance compared to dendripolylysine and enables the release of drugs at effective concentrations to obtain better therapeutic effects. In order to obtain better therapeutic effects, an effective drug concentration could be released.

[0893] Biological Safety Evaluation Example C: 101A1603 vs 101B00E SN-38 Enzyme Drug Release Rate

[0894] [Table 17]

[0895] [Table 18]

[0896] The polymer solution added with papain buffer was placed in a 37°C water bath, shaken, and the released polymer solution was taken out from the test tube at different time points, returned to room temperature, 0.500 mL was pipetted, diluted with 0.500 mL of DMSO solution, the concentration of the released SN-38 was measured by HPLC, and the release rate of SN-38 was calculated.

[0897] [Table 19]

[0898] Over 75 hours, the average drug release rate of ε-poly-L-lysine derivative-drug coupling 101B00E was 5.6 times higher than that of the dendrimer (101A1603) (Figure 3). Similarly, this result indicates that ε-poly-L-lysine derivative-drug coupling has lower site resistance and is advantageous for the release of the active ingredient of the drug and the treatment of diseases.

[0899] Biological Safety Evaluation Example D: 101C00A and 101D00A SN-38 Enzyme Drug Release Rate

[0900]

Table 20

[0901]

Table 21

[0902] The polymer solution added with papain buffer was placed in a water bath at 37 °C, shaken, and the released polymer solution was taken out from the test tube at different time points, returned to room temperature, 0.500 mL was pipetted, diluted with 0.500 mL of DMSO solution, the concentration of the released SN-38 was measured by HPLC, and the release rate of SN-38 was calculated.

[0903]

Table 22

[0904] The average drug release rate of ε-poly-L-lysine derivative-drug coupling 101C00A over 75 hours was 2.33 times higher than that of 101D00A (Figure 4).

[0905] Biological safety evaluation Example E: Comparison of plasma / tumor drug distribution in HepG2 hepatocellular carcinoma CDX model mice after intravenous injection of irinotecan against 101B00E (calculated based on 10 mg / kg of SN-38) 4 - 5-week-old, 20 - 25 g BALB / C nude mice were bred in the experimental environment for 1 week after arrival, and HepG2 tumor blocks were subcutaneously inoculated on the right back of the nude mice. When the tumor growth reached 200 - 400 mm 3 drug administration was started in the loaded nude mouse group at 3 mice / group / time point. The tumor volume was calculated as V = 0.5a × b 2 where a and b represent the major axis and minor axis of the tumor, respectively.

[0906] Irinotecan and 101B00E were both administered via the tail vein at a dose equivalent to 10 mg / kg of SN-38. The mice were sacrificed at the designated time points, and the blood was centrifuged to obtain plasma. The tumor tissues were isolated, temporarily stored in dry ice, and then transferred to an -80°C refrigerator for backup.

[0907] The tumor tissues were thawed, the tumor weights were measured, a certain amount of a 50 / 50 methanol and water mixture was added, and they were homogenized using a handheld homogenizer on an ice bath. The homogenate was centrifuged at 4°C at 6000×g for 10 minutes, and the supernatant was collected.

[0908] 30 μL of plasma or supernatant after tumor homogenization was taken, and 200 μL of acetonitrile in which the internal standard substance (toluenesulfonyl urea, 50 ng / mL) was dissolved was added. The mixture was vortexed for 5 minutes, centrifuged at 6000×g for 10 minutes, and then 75 μL of the supernatant was transferred to a 96-well plate pre-added with 75 μL of water, shaken at 500 rpm for 5 minutes, and 2 μL of LC MS / MS was injected for analysis.

[0909] Chromatography column: ACQUITY UPLC BEH C18 (2.1×50 mm, 1.7 μm).

[0910] Phase A: 0.1% formic acid / water, Phase B: 0.1% formic acid / acetonitrile.

[0911] The gradient was 90% Phase A for 0.3 minutes, 10% Phase A from 1.5 - 2.3 minutes, 10 - 90% Phase A from 2.30 - 2.31 minutes, and 90% Phase A from 2.31 - 2.8 minutes.

[0912] In the 101B00E group, 10112 was used as a standard sample to detect the conversion of 10112 released in plasma and tumors to SN-38, and in the irinotecan group, the released SN-38 was detected.

[0913]

Table 23

[0914] Refer to Figure 5 for the SN-38 tumor / blood concentration-time curve generated by the metabolism of irinotecan.

[0915]

Table 24

[0916] Refer to Figure 6 for the tumor / plasma concentration-time curve of 10112 (piperazine-10-O-SN-38) generated by the metabolism of 101B00E.

[0917]

Table 25

[0918] From the above table, it can be concluded that the tumor / plasma ratio of piperazine-10-O-SN38 (10112) released by the ε-poly-L-lysine derivative-drug coupling 101B00E reached a maximum of 21.2 ± 6.5 at 24 hours (Figure 7), while the tumor / plasma ratio of SN-38 released by irinotecan reached a maximum of 4.4 ± 1.2 at 8 hours. Compared with the conventional chemotherapeutic drug irinotecan, the ε-poly-L-lysine derivative-drug conjugate 101B00E of the invention has better tissue-specific distribution and significant targeted delivery to the tumor site.

[0919] Biological Safety Evaluation Example F: In Vivo Efficacy Comparison 1 BALB / C nude mice aged 4 - 5 weeks and weighing 20 - 25 g were bred in the experimental environment for 1 week after arrival. HepG2 tumor blocks were subcutaneously inoculated on the right back of the nude mice, and administration was started from the group where the tumor growth reached 160 mm 3 and each group consisted of 6 - 7 mice. The changes in body weight and tumor were measured twice a week, and the inhibition, delay, and cure of tumor growth were used as experimental indicators. The tumor diameter was measured twice a week with vernier calipers. The tumor volume was V = 0.5a × b2 It was calculated as follows. a and b indicate the major axis and minor axis of the tumor, respectively.

[0920] The tumor suppression effect of the compound was evaluated by the relative tumor growth rate T / C (%). The evaluation criteria were as follows: T / C (%) > 40% was regarded as ineffective, and T / C (%) ≤ 40% and those treated with statistically P < 0.05 were regarded as effective.

[0921] Relative tumor growth rate T / C (%): The calculation formula was as follows: T / C% = T RTV ÷C RTV ×100% (T RTV : The average value of RTV in the treatment group, C RTV : The average value of RTV in the negative control group). The relative tumor volume (RTV) was calculated from the measurement results of the tumor, and the formula was RTV = V t / V0. Here, V0 was the tumor volume measured at the time of drug administration to the group (i.e., d0), and V t was the tumor volume at a specific measurement time, and T RTV and C RTV were obtained from the same-day data.

[0922] Tumor suppression rate (%) = 100% - T / C (%) (1) Comparison of in vivo efficacy between ε-poly-L-lysine derivative-drug coupling 101B00A and dendritic polymer coupling 101A1601: As can be seen from Figure 8, in the CDX model of HepG2 hepatocellular carcinoma, 101B00A with much smaller enzyme attack site resistance significantly exceeded 101A1601 and doxorubicin hydrochloride in tumor suppression. However, in the CDX model of HepG2 hepatocellular carcinoma, no statistically significant difference was observed between 101A1601 and doxorubicin hydrochloride. This suggests that the ε-poly-L-lysine derivative-drug coupling 101B00A with a faster enzyme release rate in vitro has a higher tumor suppression effect in vivo than the dendritic polylysine 101A1601.

[0923] (2) Comparison of in vivo efficacy between ε-poly-L-lysine derivative-drug coupling 101B00C and ε-poly-L-lysine derivative-drug coupling 101B00E: As can be seen from Figure 9, in the HepG2 hepatocellular carcinoma CDX model, on the 33rd day, 101B00C showed a tumor suppression rate of 93% and 101B00E showed a tumor suppression rate of 92%, while the positive drug group, irinotecan, showed a tumor suppression rate of -49%. Therefore, compared with the positive drug irinotecan, both ε-poly-L-lysine derivative-drug couplings 101B00C and 101B00E had excellent tumor suppression ability.

[0924] (3) Comparison of in vivo efficacy between ε-poly-L-lysine derivative-drug coupling 101C00A and ε-poly-L-lysine derivative-drug coupling 101D00A: As can be seen from Figure 10, in the HepG2 hepatocellular carcinoma CDX model, on the 33rd day, 101C00A showed a tumor suppression rate of 90% and 101D00A showed a tumor suppression rate of 91%, while the positive drug group, irinotecan, showed a tumor suppression rate of -49%. It was shown that they had excellent tumor suppression ability.

[0925] Biological safety evaluation Example G: Comparison of the efficacy between 101B00G and nanoalbumin paclitaxel in a subcutaneous pancreatic cancer transplantation model 2 BALB / C nude mice at 4 - 5 weeks of age and weighing 20 - 25 g were bred in the experimental environment for 1 week after arrival. BxPC-3 tumor blocks were subcutaneously inoculated on the right back of the nude mice. Tumor growth reached an average volume of 157 mm 3 at the start of drug administration between groups, and 6 mice were in each group. The changes in body weight and tumors were measured twice a week, and the presence or absence of tumor growth suppression, delay, and cure were used as experimental indicators. The tumor diameter was measured twice a week with vernier calipers. The tumor volume was calculated as V = 0.5a × b 2 where a and b represent the major and minor axes of the tumor, respectively.

[0926] The tumor suppression effect of the compound was evaluated by the relative tumor growth rate T / C (%). The evaluation criteria were as follows: T / C (%) > 40% was regarded as ineffective, and T / C (%) ≤ 40% and those treated with statistically P < 0.05 were regarded as effective.

[0927] Relative tumor growth rate T / C (%): The calculation formula was as follows: T / C% = T RTV ÷C RTV ×100% (T RTV : The average value of RTV in the treatment group, C RTV : The average value of RTV in the negative control group). The relative tumor volume (RTV) was calculated from the tumor measurement results, and the formula was RTV = V t / V0. Here, V0 was the tumor volume measured at the time of drug administration to the group (i.e., d0), and V t was the tumor volume at a specific measurement time, T RTV and C RTV were obtained from the same-day data.

[0928] Tumor suppression rate (%) = 100% - T / C (%) ε-Poly-L-lysine derivative-SN-38 coupling 101B00G and nanoalbumin paclitaxel: As can be seen from Figure 11, 21 days after administration, the tumor volume of the blank control group of homozygous mice was 959 ± 113 mm 3It was reached. The nanoalbumin-paclitaxel administration group was significantly different from the blank control group (T / C = 30%, p = 0.0006), and ε-poly-L-lysine derivative-SN-38 coupling 101B00G (20 mg / kg) was significantly different from the blank control group (T / C = 5%, p = 0.00006). 101B00G (10 mg / kg) was significantly different from the blank control group (T / C = 9%, p = 0.0001), and 101B00G (5 mg / kg) was significantly different from the blank control group (T / C = 27%, p = 0.0016). This indicates that the efficacy (MTD) of paclitaxel albumin at 60 mg / kg can be achieved with 101B00G at 5 mg / kg, and further increasing 101B00G to 20 mg / kg enhances the inhibitory effect on BxPC-3 pancreatic cancer. Even when 101B00G was administered at 20 mg / kg, the impact on the body weight of mice was smaller than that when paclitaxel albumin was administered at 60 mg / kg. The effect of 101B00G on BxPC-3 pancreatic cancer was significantly superior to that of paclitaxel albumin, the first-choice drug, suggesting the possibility of clinical application.

[0929] Biological safety evaluation example H: In vivo pharmacokinetic study using 101B00G rats SD male rats weighing 220 - 250 g were bred in the experimental environment for 1 week after arrival. Six rats were used for single-dose and single-sex experiments, and blood samples were collected alternately (3 rats had blood samples collected at pre-dose, 0.25 hours, 1 hour, 4 hours, and 24 hours, and the other 3 rats had blood samples collected at 0.083 hours, 0.5 hours, 2 hours, 8 hours, 48 hours, and 72 hours).

[0930] 101B00G was dissolved in 5% glucose solution and administered via the tail vein at a dose of 15 mg / kg (SN-38) at 10 ml / kg once a week for a total of 4 times. Approximately 0.2 ml of whole blood was collected from the jugular vein into an EDTA-2K anticoagulant tube at the defined time points before and after the first and fourth administrations. After centrifugation at 6000×g for 10 minutes, the upper layer was collected. Plasma was centrifuged at 6000×g for 10 minutes, and the upper-layer plasma was collected to measure the free SN-38 concentration and the polymer-bound SN-38 concentration.

[0931] The released SN-38 concentration was measured: 30 μL of plasma was collected, and 200 μL of an acetonitrile / methanol mixed solution containing an internal standard substance (10-hydroxycamptothecin, 50 ng / mL) was added. After vortexing the mixture for 5 minutes, it was centrifuged at 6000×g for 10 minutes. 75 μL of the supernatant was transferred to a 96-well plate pre-filled with 75 μL of water, vortexed at 500 rpm for 5 minutes, and then 2 μL of the sample was injected into the LC MS / MS to analyze the release of the SN-38 concentration.

[0932] Measurement of total SN-38 concentration (polymer-bound): After 4 hours, the vial was taken out from the chromatography injection vial, cooled to room temperature, then 10 μl of 6.5M NaOH was added, capped and sealed, vortex-mixed at 1000 rpm for 30 seconds, and reacted at room temperature for 15 minutes. 480 μl of 8-fold volume of the precipitant (volume ratio of methanol to acetonitrile 1:1) was added, vortex-mixed at 1000 rpm for 5 minutes, and centrifuged at 4℃, 4700g for 10 minutes. 130 μL of the supernatant was transferred to a 96-well plate according to the plate map, 2 μl of 50% trifluoroacetic acid solution was added and mixed well. The plate was sealed with aluminum foil, transferred to the autosampler, and the concentration of the released SN-38 was measured.

[0933] The released SN-38 concentration and the total SN-38 concentration in plasma after the first dose are shown in Figure 13, the released SN-38 concentration and the total SN-38 concentration in plasma before and after the fourth dose are shown in Figure 14, and the calculated ratio of the released SN-38 concentration to the total SN-38 concentration in plasma at each time point is shown in the following two tables:

[0934]

Table 26

[0935]

Table 27

[0936] In some embodiments, as a result of intravenous administration of 101B00G 15 mg / kg (SN-38) to rats via the tail vein, the ratio of the SN-38 released into the plasma to the total SN-38 was relatively high, at 0.16 - 0.03%, only during the initial 0.083 - 2 hours after administration. From 4 hours to 168 hours after administration (0 hours on the 22nd day corresponds to 168 hours after the third administration), the released SN-38 accounted for 0.012 - 0.015% of the total SN-38, suggesting that 101B00G is extremely stable in rat plasma and does not release SN-38 non-specifically into the plasma. As can be seen from FIGS. 13 and 14, the polymer-bound SN-38 has a long half-life, indicating that 101B00G has a long circulation effect in the plasma.

[0937] Biological Safety Evaluation Example I: 1200 mm 3 Comparison of the Efficacy of 101B00G and Topotecan against the H69 Small Cell Lung Cancer Large Tumor Subcutaneous Implantation Model BALB / C nude mice, 4 - 5 weeks old and weighing 20 - 25 g, were housed in the experimental environment for 1 week after arrival. H69 small cell lung cancer tumor blocks were subcutaneously inoculated on the right back of the nude mice, and group drug administration was started when tumor growth reached ~1200 mm 3 . The changes in body weight and tumor were measured twice a week, and the inhibition, delay, and cure of tumor growth were used as experimental indicators. The tumor diameter was measured twice a week with vernier calipers. The tumor volume was calculated as V = 0.5a × b 2 . a and b represent the major axis and minor axis of the tumor, respectively.

[0938] As derived from FIG. 15, when topotecan was administered once a day for 5 days using the literature MTD dose of 2 mg / kg (converted to free base amount), the inhibitory effect on small cell lung cancer of 1200 mm 3 was low, and the tumor volume on the 22nd day increased by 22% from the initial value. In contrast, 101B00G continued to reduce the tumor volume even after a single administration, and the tumor volume on the 22nd day decreased by 81% from the initial value. 101B00G was shown to be significantly superior to existing clinical second-choice drugs for small cell lung cancer, indicating the potential for clinical application.

[0939] Example of Biological Safety Evaluation J: 600 mm 3 Efficacy of 101B00G in a Subcutaneous Large Tumor Xenograft Model of HepG2 Hepatocellular Carcinoma 4 - 5 - week - old, 20 - 25 g BALB / C nude mice were bred in the experimental environment for 1 week after arrival. HepG2 tumor blocks were subcutaneously inoculated into the right back of the nude mice, and group drug administration was started when tumor growth reached ~600 mm 3 Twice a week, body weight and tumor changes were measured, and the presence or absence of tumor growth inhibition, delay, and cure were used as experimental indicators. The tumor diameter was measured twice a week with calipers. The tumor volume was calculated as V = 0.5a×b 2 where a and b represent the major and minor diameters of the tumor, respectively.

[0940] As can be seen from Figure 16, in the 600 mm subcutaneous xenograft model of large HepG2 hepatocellular carcinoma, the tumor volume continued to decrease after administration of 101B00G, indicating the potential clinical application of 101B00G. 3 Even in the subcutaneous xenograft model of large HepG2 hepatocellular carcinoma, the tumor volume continued to decrease after administration of 101B00G, indicating the potential clinical application of 101B00G.

[0941] Example of Biological Safety Evaluation K: Comparison of the Efficacy of 101T00A - O and Albumin - Paclitaxel in a Subcutaneous Pancreatic Cancer Xenograft Model 4 - 5 - week - old, 20 - 25 g BALB / C nude mice were bred in the experimental environment for 1 week after arrival. BxPC - 3 tumor blocks were subcutaneously inoculated into the right back of the nude mice, and group drug administration was started when tumor growth reached 157 mm 3 Twice a week, body weight and tumor changes were measured, and the presence or absence of tumor growth inhibition, delay, and cure were used as experimental indicators. The tumor diameter was measured twice a week with calipers. The tumor volume was calculated as V = 0.5a×b 2 where a and b represent the major and minor diameters of the tumor, respectively.

[0942] The tumor - suppressing effect of the compound was evaluated by the relative tumor growth rate T / C (%). The evaluation criteria were as follows: T / C (%) > 40% was regarded as ineffective, and T / C (%) ≤ 40% and statistically processed with P < 0.05 was regarded as effective.

[0943] Relative tumor growth rate T / C (%): The calculation formula is as follows: T / C% = T RTV ÷C RTV ×100% (T RTV : The average value of RTV in the treatment group, C RTV : The average value of RTV in the negative control group). The relative tumor volume (RTV) was calculated from the tumor measurement results, and the formula was RTV = V t / V0. Here, V0 is the tumor volume measured at the time of drug administration to the group (i.e., d0), and V t is the tumor volume at a specific measurement time, T RTV and C RTV were obtained from the same-day data.

[0944] Tumor inhibition rate (%) = 100% - T / C (%) As can be seen from Figure 17, 21 days after administration, the tumor volume of the blank control group of homozygous mice reached 959 ± 113 mm 3 . The nanoalbumin-paclitaxel group (60 mg / kg) was significantly different from the blank control group (T / C = 30%, p = 0.0007), and 101T00A-O was significantly different from the blank control group (T / C = 7%, p = 0.00006). On the 21st day, the efficacy of 101T00A-O was significantly better than that of the paclitaxel albumin group (60 mg / kg), and there is a possibility of clinical application.

[0945] As described above, the specific embodiments of the present invention have been described, but these are merely examples, and those skilled in the art should understand that various changes or modifications can be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the protection scope of the present invention is limited by the appended claims.

Brief Description of the Drawings

[0946]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Figure 13

Figure 14

Figure 15

Figure 16

Figure 17

Claims

1. An ε-poly-L-lysine derivative-drug conjugate represented by formula (I): 【Chemical Formula 1】 The ε-poly-L-lysine derivative-drug conjugate includes the following: (1) It contains an ε-poly-L-lysine residue represented by formula (II), and the number of repeating units of the ε-poly-L-lysine body is n, 【Chemical Formula 2】 Here, the ε-poly-L-lysine is of biosynthetic origin or chemical synthetic origin; The value of n is selected from integers of 10 to 70; (2) The linkers L0 and L1 are covalent bonds, L2 is an environmentally responsive linker having a structure of L2a - L2b, L2a is linked to Y, L2b is linked to D-type, L2a or L2b exists alone or together; and L2a or L2b is each independently a covalent bond or an environmentally responsive linker; the environmentally responsive linker is one or more of an enzyme-responsive linker, a pH-responsive linker, a light-responsive linker, and a redox-responsive linker; The enzyme-responsive linker is characterized by comprising one or more of the following amino acid sequences: Cit-Phe, Lys-Lys, Phe-Lys, Arg-Arg, Val-Cit, Val-Ala, Val-Lys, Ala-Lys, Phe-Cit, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Arg, Asn-Pro-Val, Gly-Pro-Nle, Glu-Val-Cit, Glu-Val-Ala, Gly-Phe-Gly, Gly-Phe-Phe, Gly-Leu-Gly, Gly-Val-Ala, Gly-Phe-Leu-Gly, Gly-Phe-Phe-Leu, Gly-Leu-Leu-Gly, Ciy-Phe-Tyr-Ala, Gly-Phe-Gly-Phe, Ala-Gly-Val-Phe, Gly-Phe-Phe-Gly, Gly-Gly-Phe-Gly, Asp-Glu-Val-Asp, Gly-Phe-Leu-Gly-Phe, Gly-Phe-Ala-Gly-Leu-Phe, Gly-Leu-Ala-Ala-Val-Ala, Gly-Gly-Phe-Leu-Gly-Phe or Gln-Ser-Phe-Arg-Phe-Lys; The pH-responsive linker comprises one or more of the following structures: hydrazone, imine, oxime, carboxylic acid ester, thioester, sulfate ester, sulfonate ester, protonate, carbonate ester, carbamate, substituted carbamate, ketoacetal, acetal, silyl ether, phosphate ester, borate ester, phosphoramidite, or cis-aconitic acid group; The photo-responsive linker comprises one or more of the following structures: o-nitrobenzene, coumarin, benzoin, BODIPY or cyanine group; The redox-responsive linker comprises one or more of the following structures: thioketone, phenylborate, phenylboronic acid, oxalate, vinyl ether, thioether group, aminoacrylate, disulfide, diselenide, 2,4-dinitrobenzenesulfonate, 2-azidomethylbenzoate, 4-azidobenzyl, unsaturated acid ester, or azobenzene group; (3) Y is at least a trifunctional branching center; when the electrophilic group of the Y molecule is bonded to L2 and L2 = L2b, L2a does not exist, and the bonds of the trifunctional branching center Y to the linkers L0, L1, and L2b are selected from any of the following structures: [Chemical Formula 3] Alternatively, when the nucleophilic group of the Y molecule is bonded alone to L2, L2a, or together with L2a and L2b, in this case, L2 = L2a or L2 = L2a - L2b, and the bonds of the trifunctional branching center Y to the linkers L0, L1, L2a, and L2b are selected from any of the following structures: [Chemical Formula 4] [Chemical Formula] [Chemical Formula] [Chemical Formula] [Chemical Formula] (4) P is a pharmacokinetic regulator residue; the pharmacokinetic regulator residue is selected from polyethylene glycol derivative residues having a repeating unit number a and a terminal group Rb; the polyethylene glycol derivative residue is selected from the following structures; [Chemical Formula 5] Here, a is selected from integers from 5 to 150, r is selected from integers from 0 to 8, and Rb is a C1 - C10 alkyl group; (5) D is a pharmaceutically active agent residue; the pharmaceutically active agent residue is an antitumor drug residue, The anti-tumor drug is selected from one or more of the following: abemaciclib, abiraterone, abrocitinib, acalabrutinib, afatinib, aldesleukin, alectinib, alflutamide, almonertinib, altretamine, amcenestrant, aminoglutethimide, amsacrine, anastrozole, anlotinib, apalutamide, apatinib, arzoxifene, asiminib, asparaginase, avapritinib, avitinib, axitinib, azacitidine, baricitinib, belinostat, bendamustine, bexarotene, bicalutamide, bicyclol, binimetinib, bleomycin, boanmycin, bortezomib, bosutinib, brigatinib, buserelin, busulfan, cabazitaxel, cabozantinib, calaspargase, calicheamycin, capecitabine, capmatinib, carboplatin, carfilzomib, carmustine, carmofur, cedazuridine, ceritinib, cetrorelix, chidamide, chlorambucil, cisplatin, cladribine, clofarabine, cobimetinib, colchicine, copanlisib, crizotinib, cyclophosphamide, cytarabine, dabrafenib, dacarbazine, dacomitinib, dactinomycin, dalpiciclib, darolutamide, dasatinib, daunorubicin, decitabine, degarelix, delgociclid, denileukin, deruxtecan, docetaxel, donafenib, doxorubicin, duvelisib, enasidenib, encorafenib, ensartinib, entrectinib, enzalutamide, enzastaurinelacestrant, epirubicin, erdafitinib, eribulin, erlotinib, estradiol, estramustine, etoposide, everolimus, exemestane, fasudil, fedratinib, filgotinib, floxuridine, fludarabine, flumatinib, fluorouracil, flutamide, fluzoparib, formestane, fostamatinib, fruquintinib, fulvestrant, gefitinib, gemcitabine, gilteritinib, giredestrant, glasdegib, goserelin, histrelin, hydroxyurea, ibrutinib, ibudilast, icariin, icotinib, idarubicin, idelalisib, ifosfamide, imatinib, imiquimod, infgratini, ingenol mebutate, interferon alpha-2b, irinotecan,ivosidenib, ixabepilone, ixazomib, lanreotide, lapatinib, larotrectinib, lenalidomide, lenvatinib, letrozole, leucovorin, leuprorelide, lomustine, lonafamb, lorlatinib, lurbinectedin, maytansine, mechlorethamine, medroxyprogesterone, megestrol, melphalan, melphalan flufenamide, mercaptopurine, methotrexate, methoxsalen, methylprednisolone, midostaurin, mitomycin, mitotane, mitoxantrone, mitozolomide, mobocertinib, monomethyl auristatin E, monomethyl auristatinF, nelarabine, nandrolone, neratinib, nearsudil, nilotinib, nilutamide, nintedanib, niraparib, octreotide, olaparib, olmutinib, omacetaxine, orelabrutinib, osimertinib, oxaliplatin, paclitaxel, pacritinib, palbociclib, pamidronate, pamparib, panobinostat, pazopanib, peficitinib, pegaptanib, pegaspargase, peg interferon alfa-2b, pemigatinib, pemetrexed, pentetreotide, pentostatin, pexidartinib, phenoxybenzamine, pidotimod, plinabulin, plitidepsin, pomalidomide, ponatinib, porfimer, pralatrexate, pralsetinib, prednisone, procarbazine, pyrotinib, quizartinib, radotinib, raloxifene, raltitrexed, regorafenib, ribociclib, rintatolimod, ripretinib, romidepsin, rucaparib, ruxolitinib, savolitinib, selinexor, selpercatinib, selumetinib, sonidegib, sorafenib, sotorasib, streptozocin, sunitinib, surufatinib, talazoparib, tamoxifen, tazemetostat, tegafur, temozolomide, temsirolimus, teniposide, tepotinib, teprenone, thalidomide, thioguanine, thiotepa, thyrotropinalpha, tipiracil, tipifarnib, tirabrutinib, tirbanibulin, tivozanib, trametinib, tofacitinib, topotecan, toremifene, trabectedin, iretinoin, trifluridine, trilaciclib, triptorelin, lucatinibupadacitinib, umbralisib, utidelone, uroacitide, valrubicin, vandetanib, yemurafenib, venetoclax, vinblastine, vincristine, vindesine, vinflunine, Vinorelbine, visimodegib, vorinostat, zanubrutinib, zoledronic acid, amatoxins, anthracyclines, anthracenes, anthramycins, auristatins, bryostatins, camptothecins, carmaphycins, combretastatin, cyclosporines, cryptomycins, ecteinascidins, ellipticenes, esperamicins, mustines, neothramycins, ozogamicins, phenoxazines, podophyllotoxins, pyrrolobenzodiazepines, sibiromycins, thailandstatins, tomamycns, tubulysins, taxanes, vinca alkaloids, 7-epipaclitaxel, 2'-acetylpaclitaxel, 10-diacetylpaclitaxel, 10-deacetyl-7-epipaclitaxel, 7-xylosylpaclitaxel, 10-deacetyl-7-glutarylpaclitaxel, 7-N,N-dimethylglycylpaclitaxel, 7-L-alanyl paclitaxel, lerontaxine, comedo, 9-aminocomedo, 9-nitocomedo, leptotecan, gimatecon, velotecan, 10-hydroxycamptothecin, 10-hydroxy-ethyl-camptothecin (SN-38), irinotecan, pimaricin, aclarubicin, sirolimus, tacrolimus, luteinizing hormone, estrogen, rapamycin, pucamycin, tritiated alkaloid or curcumin (6) X is a terminal group, and the terminal group is selected from OR, SR, NR1R2, a carboxyl protecting group, or L2b-D, where R, R1, and R2 are independently selected from H, a C1-C30 alkyl group, a C1-C10 alkyl group substituted with R1-1, a C1-C30 alkoxy group, a C3-C30 alkenyl group, a C3-C30 alkynyl group, a C3-C8 cycloalkyl group, a C2-C8 heterocycloalkyl group, a C6-C20 aryl group, or a C5-C20 heteroaryl group; alternatively, R1, R2, and the linked N atom form a C2-C8 heterocycloalkyl group; the heteroatom in the C2-C8 heterocycloalkyl group is O, S, or N; the number of heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different: the heteroatom in the C5-C20 heteroaryl group is O, S, or N, the number of heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different; R1-1 is -NH2 or [Chemical Formula 6] and is.

2. The ε-poly-L-lysine derivative-drug conjugate according to Claim 1, wherein the value of the number of repeating units n of the ε-poly-L-lysine moiety is an integer from 20 to 40.

3. The ε-poly-L-lysine derivative-drug conjugate according to Claim 2, wherein the value of the number of repeating units n of the ε-poly-L-lysine moiety is an integer of 30.

4. The ε-poly-L-lysine derivative-drug conjugate according to Claim 1, wherein the structure of the ε-poly-L-lysine derivative-drug conjugate is as shown in formula (IV): [Chemical Formula 7] Here, the value of n is an integer from 10 to 40, and X, Y, P, and D are as defined in Claim 1, respectively.

5. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein when the electrophilic group of the Y molecule is bonded to L ２ and L ２ = L ２ｂ exists, L ２ａ does not exist, and the bonds of the linker L ０ , L １ and L ２ｂ to the trifunctional branching center Y are selected from any of the following structures: 【Chemical formula 8】 Alternatively, when the nucleophilic group of the Y molecule is bonded to L ２ , L ２ａ alone, or L ２ａ and L ２ｂ are bonded together, in this case, L ２ = L ２ａ or L ２ = L ２ａ -L ２ｂ and the bonds of the linker L ０ , L １ , L ２ａ and L ２ｂ to the trifunctional branching center Y are selected from any of the following structures: 【Chemical formula 9】 .

6. The ε-poly-L-lysine derivative-drug conjugate according to claim 5, wherein when the electrophilic group of the Y molecule is bonded to L2, and L2 = L2b, L2a does not exist, and the bonds of the linker L0, L1 and L2b to the trifunctional branching center Y are selected from any of the following structures: 【Chemical formula 10】 .

7. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the terminal group X is OR, SR, NR １ R ２ , a carboxy protecting group or -L ２ｂ -D, and the R, R １ , R ２ are independently H, C １ ~C １０ an alkyl group or R １－１ substituted C １ ~C １０ selected from alkyl groups. **Claim 8**: The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the enzyme-responsive linker is cleavable by one or more of the following enzymes: secreted phospholipase A2, acid phosphatase, serum alkaline phosphatase, cytochrome P450, sulfatase, prostate-specific antigen, phospholipase A1, phospholipase A2, phospholipase B, phosphodiesterase C, phosphodiesterase D type, neutrophil elastase, cysteine protease-3, histone protease, matrix metalloprotease, β-glucosidase, β-galactosidase, DTP, nitroreductase, reduced coenzyme II, aminopeptidase N, carboxylesterase, myocardial xanthine oxidase, histone deacetylase, asparagin endopeptidase, urokinase-type fibrinogen activator, urokinase-type fibrinogen activator receptor, collagenase; preferably, cleaved by one or more of the following enzymes: cysteine protease-3, histone protease, matrix metalloprotease, β-glucosidase. **Claim 9** The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the ε-poly-L-lysine derivative-drug conjugate satisfies one or more of the following conditions: (1) The enzyme-responsive linker contains the following structure: [Chemical formula 11] [Chemical formula] [Chemical formula] (2) The pH-responsive linker contains the following structure: [Chemical formula 12] where m = 0 to 4; (3) The photo-responsive linker contains the following structure: [Chemical formula 13] (4) The redox-responsive linker includes the following structure: [Chemical Formula 14] .

10. The linker L ２ａ is a covalent bond or a linker represented by formula (V), and the ε-poly-L-lysine derivative-drug conjugate according to claim 1 is characterized in that: [Chemical Formula 15] The Q is selected from any of the following; [Chemical Formula 16] Here, R ３ and R ４ are independently H, D, C １ to C ６ alkyl group, C １ to C ６ alkoxy group, C ３ to C ６ alkenyl group, C ３ to C ６ alkynyl group, C ３ to C ８ cycloalkyl group, C ２ to C ８ heterocycloalkyl group, C ６ to C １０ aryl group, or C ５ to C １０ heteroaryl group; or here, R3, R4 and the linked C atoms form a C ３ to C ８ alkyl group or heterocycloalkyl group; the heteroatom in the C ２ to C ８ heterocycloalkyl group is O, S or N, the number of the heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different; the heteroatom in the C ５ to C １０ heteroaryl group is O, S or N, the number of the heteroatoms is one or more, and in the case of one or more, the heteroatoms are the same or different; W is a covalent bond or C ０ -C ２０ ; a fragment that may or may not contain a heteroatom, where the heteroatom is O, S, Se, N, P, Si, or B, the number of said heteroatoms is one or more, and in the case of one or more, the said heteroatoms are the same or different; whether the W fragment contains an unsaturated bond or not; Z is 【Chemical Formula 17】 selected from the species of, where R ａ is H, C １ -C １０ alkyl group, C １ -C １０ alkoxy group, C ３ -C １０ alkenyl group, C ３ -C １０ alkynyl group, C ３ -C ８ cycloalkyl group, C ２ -C ８ heterocycloalkyl group, C ６ -C １０ aryl group or C ５ -C １０ heteroaryl group; the heteroatom in the C ２ -C ８ heterocycloalkyl group is O, S or N, the number of said heteroatoms is one or more, and in the case of one or more, the said heteroatoms are the same or different; the heteroatom in the C ５ -C １０ heteroaryl group is O, S or N, the number of said heteroatoms is one or more, and in the case of one or more, the said heteroatoms are the same or different.

11. The linker L ２ａ is selected from the following structures or a covalent bond, and the ε-poly-L-lysine derivative-drug conjugate according to claim 10 is characterized in that: 【Chemical Formula 18】 where, A is O, S, S(O), S(O) ２ , NR a, C(R ３ R ４ ) is; p is selected from integers from 0 to 16; g is selected from integers from 0 to 16; m is selected from integers from 0 to 4; R ａ is the same as the definition described in claim 10; R ３ and R ４ are the same as the definition described in claim 10.

12. The linker L ２ａ is selected from the following structures, and the ε-poly-L-lysine derivative-drug conjugate according to claim 1: 【Chemical Formula 19】 .

13. The linker L ２ｂ is selected from the following structures, and the ε-poly-L-lysine derivative-drug conjugate according to claim 1: 【Chemical Formula 20】 .

14. The linker L ２ｂ is selected from the following structures, and the ε-poly-L-lysine derivative-drug conjugate according to claim 1: 【Chemical Formula 21】 .

15. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the pharmacokinetic regulator residue P satisfies one or more of the following conditions: (1) a is an integer from 10 to 60; (2) R ｂ is an alkyl group of C １ to C ６ .

16. An ε-poly-L-lysine derivative-drug conjugate according to claim 15, wherein the pharmacokinetic regulator residue P satisfies one or more of the following conditions: (1) a is an integer from 15 to 50; (2) Rb is an alkyl group having 1 to 3 carbon atoms.

17. An ε-poly-L-lysine derivative-drug conjugate according to claim 15, wherein the pharmacokinetic regulator residue P satisfies one or more of the following conditions: (1) a is an integer of 21, 43 or 44; (2) Rb is a methyl group, an ethyl group, n-propyl or isopropyl.

18. An ε-poly-L-lysine derivative-drug conjugate according to claim 15, wherein the pharmacokinetic regulator residue P is 【Chemical formula 22】 where a is selected from integers from 20 to 45, and r is selected from integers from 0 to 3.

19. An ε-poly-L-lysine derivative-drug conjugate according to claim 18, wherein the pharmacokinetic regulator residue P is a methyl-capped polyethylene glycol derivative residue of the number of repeating units a, and the methyl-capped polyethylene glycol derivative residue is selected from the following structures: 【Chemical formula 23】 Here, a is selected from integers from 5 to 150, and r is selected from integers from 0 to 8.

20. The methyl-capped polyethylene glycol derivative residue is 【Chemical formula 24】 The ε-poly-L-lysine derivative-drug conjugate according to claim 19, characterized in that it is as follows.

21. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the ε-poly-L-lysine derivative-drug conjugate satisfies one of the following conditions: (1) The anti-tumor drug is aldesleukin, abemaciclib, abiraterone, abrocitinib, acalabrutinib, afatinib, alectinib, alflutamide, almonertinib, amcenestrant, anastrozole, anlotinib, apalutamide, apatinib, arzoxifene, asiminib, avapritinib, avitinib, axitinib, baricitinib, belinostat, bexarotene, bicalutamide, binimetinib, bleomycin, boanmycin, bortezomib, bosutinib, brigatinib, buserelin, cabozantinib, capmatinib, carfilzomib, carmustine, ceritinib, cetrorelix, chidamide, cobimetinib, copanlisib, crizotinib, dabrafenib, dacomitinib, dalpicilib, darolutamide, dasatinib, degarelix, delgociclib, donafenib, duvelisib, enasidenib, encorafenib, ensartinib, entrectinib, enzalutamide, enzastaurin, elacestrant, erdafitinib, erlotinib, everolimus, fedatinib, filgotinib, flumatinib, fluzoparib, formestane, fostamatinib, fnquintinib, fulvestrant, gefitinib, gilteritinib, giredestrant, glasdegib, goserelin, histrelin, ibrutinib, ibudilast, icotinib, idarubicin, idelalisib, imatinib, imiquimod, infgratiniib,ivosidenib, ixazomib, lanreotide, lapatinib, larotrectinib, lenalidomide, lenvatinib, letrozole,It is an oncology target drug selected from one or more of leucovorin, leuprolide, lonafarnib, lorlatinib, medroxyprogesterone, megestrol, methylprednisolone, midostaurin, mobocertinib, nandrolone, neratinib, nilotinib, nilutamide, nintedanib, niraparib, olaparib, olmutinib, orelabrutinib, osimertinib, pacritinib, palbociclib, pamidronate, pamparib, panobinostat, pazopanib, peficitinib, pegaptanib, pemigatinib, pexidartinib, pidotimod, pomalidomide, ponatinib, prasetinib, pyrotinib, quizartinib, radotinib, raloxifene, regorafenib, ribociclib, rintatolimod, ripretinib, rucaparib, ruxolitinib, savolitinib, selinexor, selpercatinib, selumetinib, sonidegib, sorafenib, sotorasib, sunitinib, surufatinib, talazoparib, tamoxifen, tazemetostat, temsirolimus, tepotinib, thalidomide, tipifamb, tirabrutinib, tivozanib, trametinib, tofacitinib, toremifene, tretinoin, trilaciclib, triptorelin, tucatinib, upadacitinib, umbralisib, vandetanib, vemurafenib, venetoclax, visimodgib, vorinostat, zanubrutinib or zoledronic acid; (2) The anti-tumor drug is altretamine, aminoglutethimide, amsacrine, asparaginase, azacitidine, bendamustine, bexarotene, bicyclol, bleomycin, boanmycin, buserelin, busulfan, cabazitaxel, calaspargase, calicheamycin, capecitabine, carboplatin, carmustine, carmofur, cedazuidine, chlorambucil, cisplatin, cladribine, clofarabine, colchicine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, decitabine, denileukin, deruxtecan, docetaxel, doxorubicin, epirubicin, eribulin, estradiol, estramustine, etoposide, exemestane, fasudil, floxuridine, fludarabine, fluorouracil, flutamide, formestane, gemcitabine, hydroxyurea, icariin, idarubicin, ifosfamide, ingenol mebutate, irinotecan, ixabepilone, leucovorin, lomustine, lurbinectedin, maytansine, mechlorethamine, medroxyprogesterone, megestrol, melphalan, melphlan flufenamide, mercaptopurine, methotrexate, methoxsalen, methylprednisolone, mitomycin, mitotane, mitoxantrone, mitozolomide, monomethyl auristatin E, monomethyl auristatinF, nelarabine, nandrolone, nearsudil, octreotide, omacetaxine, oxaliplatin, paclitaxel, pamidronate, pemetrexed, pentetreotide, pentostatin, phenoxybenzamine, plinabulin, plitidepsin, porfimer, pralatrexate, prednisone, procarbazine, procarbazine, raltitrexed, romidepsin, streptozocin, tegafur, temozolomide, teniposide, teprenone, thioguanine, thiotepa, thyrotropin alpha, tipiracil, tirbanibulin, topotecan, trabecetedine, trifluridine, utidelone, uroacitide, valrubicin, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, amatoxins, anthracyclines, anthracenes, anthramycins, auristatins, bryostatins, camptothecins, camaphycins, combretastatin, cyclosporines, cryptomycins, ecteinascidins, ellipticenes, esperamicins, mustines, neothramycins, ozogamicins, phenoxazines, podophyllotoxins, pyrrolobenzodiazepines, sibiromycins, thailandstatins, tomamycns, tubulysins, taxanes, vincaOne or more chemotherapeutic agents selected from alkaloids, 7-epipaclitaxel, 2'-acetylpaclitaxel, 10-diacetylpaclitaxel, 10-deacetyl-7-epipaclitaxel, 7-xylosylpaclitaxel, 10-deacetyl-7-glutaryl paclitaxel, 7-N,N-dimethylglycylpaclitaxel, 7-L-alanyl paclitaxel, leurotaxine, comedo, 9-aminocomedo, 9-nitromedo, camptothecin, gimatecon, velotecan, 10-hydroxycamptothecin, 10-hydroxy-ethyl-camptothecin (SN-38), irinotecan, pimaricin, aclarubicin, sirolimus, tacrolimus, luteinizing hormone, estrogen, rapamycin, pucamycin, tritiated alkaloid or curcumin.

22. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the pharmaceutical active residue D is selected from one of the following structures: 【Chemical Formula 25】 【Chemical Formula】 【Chemical Formula】 。

23. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the ε-poly-L-lysine derivative-drug conjugate is selected from one of the following structures: 【Chemical Formula 26】 【Chemical Formula】 【Chemical Formula】 【Chemical Formula】 Here, L ２ａ 、L ２ｂ each has the same definition as in claim 1; D has the same definition as in claim 1; X has the same definition as in claim 1; R ｂ has the same definition as in claim 1; n is an integer from 10 to 70; a is an integer from 5 to 150.

24. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the ε-poly-L-lysine derivative-drug conjugate is selected from one of the following structures: 【Chemical Formula 27】 Here, L ２ａ and L ２ｂ are each the same as defined in claim 1; D is the same as defined in claim 1: X is the same as defined in claim 1; R ｂ is the same as defined in claim 1; n is an integer from 10 to 70; a is an integer from 5 to 150.

25. R ｂ is a methyl group, and the number average molecular weight of the polyethylene glycol derivative residue is 550, 1,000, 2,000, 3,000, 4,000 or 5,000, and the ε-poly-L-lysine derivative-drug conjugate according to claim 1.

26. X is -OH, 【Chemical Formula 28】 or -L ２ｂ -D; the -L ２ｂ -D is selected from one of the following structures, and the ε-poly-L-lysine derivative-drug conjugate according to claim 1: 【Chemical Formula 29】 【Chemical Structure】 【Chemical Structure】 。

27. L ２ｂThe ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein it is the enzyme-responsive linker or the pH-responsive linker.

28. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein D is a residue of a chemotherapeutic agent, and the type of the chemotherapeutic agent is selected from one or more of amatoxins, anthracyclines, anthracenes, anthramycins, auristatins, bryostatins, camptothecins, carmofur, combretastatin, cyclosporins, cryptomycins, ecteinascidins, ellipticines, esperamicins, maytansines, mustines, neothramycins, ozogamicins, phenoxazines, platinum complexes, podophyllotoxins, pyrrolobenzodiazepines, sibiromycins, tyranstatin, tomatamycin, tubulysins, taxanes or vinca alkaloids.

29. The ε-poly-L-lysine derivative-drug conjugate according to claim 28, wherein the chemotherapeutic drug is selected from bleomycin, boanamycin, cabazitaxel, calicheamicin, carboplatin, cisplatin, dactinomycin, daunorubicin, deruxtecan, docetaxel, doxorubicin, epirubicin, eribulin, etoposide, idarubicin, irinotecan, ixabepilone, lurbinectedin, maytansinol, monomethyl auristatin E, monomethyl auristatin F, mitomycin, oxaliplatin, paclitaxel, streptozocin, teniposide, topotecan, trabectedin, valrubicin, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, 7-epipaclitaxel, 2'-acetylpaclitaxel, 10-deacetylpaclitaxel, 10-deacetyl-7-epipaclitaxel, 7-xylosylpaclitaxel, 10-deacetyl-7-glutaryl paclitaxel, 7-N,N-dimethylglycylpaclitaxel, 7-L-alanyl paclitaxel, lerotaxectin, comedocarcin, 9-aminocamptothecin, 9-nitrocamptothecin, leptotecan, gimatecan, vertotecan, 10-hydroxycamptothecin, 10-hydroxy-7-ethylcamptothecin (SN-38), exatecan, vibramycin, aclacinomycin, or procarbamptothecin.

30. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the ε-poly-L-lysine derivative-drug conjugate satisfies one or more of the following conditions: (1) The branching center Y is linked to the ε-poly-L-lysine residue by a linker L ０ and the molar ratio of the branching center Y to the ε-poly-L-lysine structural unit is 0.5:1 to 1.5:1; (2) The pharmacokinetic regulator residue P is linked to the branching center Y via a linker L １ and the molar ratio of the pharmacokinetic regulator residue to the ε-poly-L-lysine structural unit is 0.5:1 to 1.5:1; (3) The pharmaceutically active agent residue D is a linker L ２Linked to the trifunctional branching center Y via, the molar ratio of the pharmaceutically active agent residue D to the ε-poly-L-lysine structural unit is 0.5:1 to 1.5:

1.

31. The ε-poly-L-lysine derivative-drug conjugate according to claim 30, characterized in that the ε-poly-L-lysine derivative-drug conjugate satisfies one or more of the following conditions: (1) The branching center Y is a linker L ０ Linked to the ε-poly-L-lysine residue via, the molar ratio of the branching center Y to the ε-poly-L-lysine structural unit is 0.8:1 to 1.2:1; (2) The pharmacokinetic regulator residue P is a linker L １ Linked to the trifunctional branching center Y via, the molar ratio of the pharmacokinetic regulator residue to the ε-poly-L-lysine structural unit is 0.8:1 to 1.2:1; (3) The pharmaceutically active agent residue D is a linker L ２ Linked to the trifunctional branching center Y via, the molar ratio of the pharmaceutically active agent residue D to the ε-poly-L-lysine structural unit is 0.8:1 to 1.2:

1.

32. The ε-poly-L-lysine derivative-drug conjugate according to any one of claims 1 to 30, characterized in that the ε-poly-L-lysine derivative-drug conjugate has an application in the prevention and treatment of tumor diseases.

33. The ε-poly-L-lysine derivative-drug conjugate according to claim 32, wherein the use of the ε-poly-L-lysine derivative-drug conjugate is for the prevention and treatment of cancer, and the cancer includes breast cancer, ovarian cancer, prostate cancer, melanoma, brain tumor, nasopharyngeal cancer, esophageal cancer, gastric cancer, hepatocellular carcinoma, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, skin cancer, glioblastoma, neuroblastoma, sarcoma, liposarcoma, osteosarcoma, bone cancer, osteosarcoma, testicular tumor, testicular tumor, uterine tumor, head and neck tumor, multiple myeloma, malignant lymphoma, polycythemia vera, leukemia, thyroid tumor, ureteral tumor, bladder tumor, gallbladder cancer, bile duct cancer, choriocarcinoma, or pediatric tumor.

34. 【Chemical formula 30】 The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein [chemical formula 31] is selected from the following structures: [Chemical formula 31] 。

35. 【Chemical formula 32】 The ε-poly-L-lysine derivative-drug conjugate according to claim 34, wherein [chemical formula 33] is selected from the following structures: [Chemical formula 33] [Chemical formula] 。

36. L ２ The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein L-D is one of the following structures: [Chemical formula 34] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] 。 Item 37: The ε-poly-L-lysine derivative-drug conjugate according to Item 36, characterized in that L2-D is one of the following structures: [Chemical Formula 35] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] 。

38. The ε-poly-L-lysine derivative-drug conjugate according to claim 1, wherein the ε-poly-L-lysine derivative-drug conjugate is selected from one of the following structures: [Chemical Formula 36] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] [Chemistry] 。

39. A compound represented by one of the formulas (VI-XV): [Chemistry 37] Here, P ０ , P ５ , P ６ are each independently selected from an OH or carboxyl protecting group, and when P ０ , P ５ , P ６ is a carboxyl protecting group, they are the same or different; P １ , P ２ , P ３ and P ４ are each independently selected from an H or amine protecting group, and when P １ , P ２ , P ３ and P ４ is an amine protecting group, they are the same or different; L １ is the same as defined in Claim 1; L ２ａ is the same as the definition recited in claim 1; P is the same as the definition recited in claim 1; n is an integer from 10 to 70; c is an integer from 0 to 3.

40. A compound according to claim 39, characterized in that the compound has the following; n is an integer from 10 to 70; L １ is a covalent bond; P is a methyl-capped polyethanol residue having a number average molecular weight of 500 to 5,000; L ２ａ is C(O)-E-C(O), where E is a covalent bond, CH ２ CH ２ -CH ２ CH ２ -CH ２ -CH ２ CH ２ -O-CH ２ CH ２ -S-CH ２ CH ２ -NH-CH ２ CH ２ -N(Me)-CH ２ or CH-CH.

41. The compound according to claim 40, characterized in that the compound is selected from one of the following compounds: 【Chemical Formula 38】 【Chemical Formula】 【Chemical Formula】 【Chemical Formula】 .

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